專利名稱：脅迫抗性植物的制作方法
技術(shù)領(lǐng)域：
(本發(fā)明)通過在植物中表達(dá)參與NAD補(bǔ)救合成途徑或NAD從頭合成途徑中的酶，提供了增加植物或植物細(xì)胞脅迫抗性的方法。
背景技術(shù)：
考慮到對(duì)最終增加作物植物的實(shí)際產(chǎn)率的永無止境的需求，對(duì)不利生長條件的耐受性是這些作物非常理想的特性，所述不利生長條件包括干旱、高光照強(qiáng)度、高溫、營養(yǎng)限制、鹽生長條件等等。
達(dá)到提高通常稱為植物脅迫抗性或脅迫耐受性目的的多種方法已有描述。由于不同的非生物脅迫條件經(jīng)常導(dǎo)致產(chǎn)生有害的活性氧(ROS))，如超氧化物或過氧化氫，因此，提高植物耐受性的初期嘗試集中在阻止產(chǎn)生ROS或?qū)⑵涑ァ＿@些方法的實(shí)施例是過表達(dá)ROS清除酶，如過氧化氫酶、過氧化物酶、超氧化物歧化酶等，或者甚至增加ROS清除分子，如抗壞血酸、谷胱甘肽等的量。產(chǎn)生脅迫耐受性植物的這些方法和其它嘗試綜述于，如wang等人，2003，Planta 2181-14。
如WO00/04173和PCT/EP2004/003995中分別描述的，植物細(xì)胞和植物的脅迫耐受性也可以通過減少內(nèi)源聚ADP核糖聚合酶(ParP)或聚(ADP-核糖)糖苷水解酶(ParG)的活性或水平而實(shí)現(xiàn)。一般認(rèn)為，在這種方法中，遭受脅迫條件的植物細(xì)胞中，導(dǎo)致創(chuàng)傷性細(xì)胞死亡的致死的NAD和ATP衰竭可以被避免或充分地推遲，以使受脅迫的細(xì)胞存活并適應(yīng)脅迫條件。
Uchimiya等人(2002)描述了命名為YK1的稻基因的分離，以及嵌合YK1基因增加含有這種基因的轉(zhuǎn)基因稻類植物對(duì)稻瘟病和幾種非生物脅迫(如NaCl、UV-C、浸沒和過氧化氫)耐受性的用途(Uchimiya等人.，2002，molecular breeding 925-31)。
Uchimiya等人還發(fā)表一篇會(huì)議摘要，描述的是在稻細(xì)胞中過表達(dá)NAD依賴性還原酶基因(YK1)還通過上調(diào)NAD合成酶的活性提高NAD(P)(H)的水平，并且推斷這種改變能依次產(chǎn)生ROS脅迫抗性所需要的氧化還原物質(zhì)庫(Uchimiya等人.2003 Keystone symposium on PlantbiologyFunctions and control of cell death，Snowbird Utah April 10-15，2003).
酵母中的NAD合成酶已經(jīng)得到很好的表征，而且它是NAD從頭合成途徑以及NAD補(bǔ)救途徑中最后的一個(gè)酶(見


圖1)。在從頭合成途徑中，喹啉(quinolate)是合成NAD的前體，并且是作為色氨酸降解產(chǎn)物產(chǎn)生。在補(bǔ)救途徑中，煙酰胺是前體分子，其為通過多種酶(如PARP、NAD依賴的脫乙酰酶或其它NAD糖苷水解酶)作用產(chǎn)生的NAD的降解產(chǎn)物。在第一步中，煙酰胺由煙酰胺酶脫去酰胺基變成煙酸。煙酸在煙酸酯磷酸核糖基轉(zhuǎn)移酶的作用下轉(zhuǎn)移到5-磷酸核糖基-1-焦磷酸上產(chǎn)生煙酸單核苷酸。這一化合物是NAD從頭合成途徑和補(bǔ)救合成途徑共同擁有的。因此，在從頭合成途徑中通過NAD+焦磷酸化酶和NAD合成酶完成這一化合物的進(jìn)一步轉(zhuǎn)化。
在酵母中，已經(jīng)將過表達(dá)PNC1(編碼煙酰胺酶)通過卡路里限制和低強(qiáng)度脅迫與生命期延長相關(guān)聯(lián)(Anderson等人，2003 Nature 423p181-185；Gallo等人，2004，Molecular and Cellular Biology 241301-1312).
對(duì)植物中NAD生物合成途徑中的各個(gè)酶所知甚少。Hunt等人2004年描述了使用可獲得的擬南芥基因組信息鑒定這些酶的植物同源物(Hunt等人，2004，New Phytologist163(1)31-44)。已經(jīng)識(shí)別的DNA序列有以下的登錄號(hào)煙酰胺酶At5g23220、At5g23230和At3g16190；煙酸酯磷酸核糖基轉(zhuǎn)移酶At4g36940、At2g23420；煙酸單核苷酸腺苷轉(zhuǎn)移酶At5g55810以及NAD合成酶At1g55090(所有的核苷酸序列在此引用作為參考)。
依然需要能增加植物脅迫抗性的備選方法，下面描述的實(shí)施方案，包括權(quán)利要求，提供了這樣的方法和手段。
發(fā)明概述在本發(fā)明的一個(gè)實(shí)施方案中，提供了獲得具有增加的脅迫抗性植物的方法，所述方法包括將嵌合基因引入植物細(xì)胞中獲得轉(zhuǎn)基因細(xì)胞，所述嵌合基因包括以下有效連接的DNA片段i.可在植物中表達(dá)的啟動(dòng)子；ii.編碼煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中植物功能酶的DNA區(qū)，所述酶選自煙酰胺酶、煙酸酯磷酸核糖基轉(zhuǎn)移酶，煙酸單核苷酸腺苷轉(zhuǎn)移酶或煙酰胺腺嘌呤二核苷酸合成酶；iii.參與轉(zhuǎn)錄終止和多聚腺苷化作用的3’末端區(qū)。
隨后，由轉(zhuǎn)基因植物細(xì)胞再生獲得轉(zhuǎn)基因植物群；并從轉(zhuǎn)基因植物群選擇比類似的非轉(zhuǎn)基因植物表現(xiàn)出增加的脅迫抗性的植物，或在脅迫條件下表現(xiàn)出降低水平活性氧的植物或維持高水平NADH的植物。DNA區(qū)可以編碼含有選自SEQ ID No.2、SEQ ID No.4、SEQ ID No.6、SEQ IDNo.8、SEQ ID No.10、SEQ ID No.12、SEQ ID No.14、SEQ ID No.16、SEQ ID No.18、SEQ ID No.20、SEQ ID No.22、SEQ ID No.24氨基酸序列的蛋白質(zhì)；或編碼具有約60％序列同一性并且具有煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的酶活性的蛋白質(zhì)，如核苷酸序列SEQ ID No.1、SEQ ID No.3、SEQ ID No.5、SEQ ID No.7、SEQ ID No.9、SEQ IDNo.11、SEQ ID No.13、SEQ ID No.15、SEQ ID No.17、SEQ ID No.19、SEQ ID No.21、或SEQ ID No.23。
在另一實(shí)施方案中，本發(fā)明涉及這里所述的嵌合基因、包含這些嵌合基因的植物細(xì)胞、基本上由含有這些嵌合基因的植物細(xì)胞組成的植物以及這樣的植物的種子。這些植物和植物細(xì)胞的特征在于其在脅迫條件下比類似的不包含這樣的嵌合基因的植物擁有更低水平的活性氧。
而在另一個(gè)實(shí)施方案中，本發(fā)明涉及使用所述的嵌合基因以增加植物的脅迫抗性，或減少在脅迫條件下植物或植物細(xì)胞中活性氧的水平。
本發(fā)明還提供編碼煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的植物功能酶的DNA序列的用途(所述酶選自煙酰胺酶、煙酸酯磷酸核糖基轉(zhuǎn)移酶、煙酸單核苷酸腺苷轉(zhuǎn)移酶或煙酰胺腺嘌呤二核苷酸合成酶)，例如編碼含有選自SEQ ID No.2、SEQ ID No.4、SEQ ID No.6、SEQ ID No.8、SEQID No.10、SEQ ID No.12、SEQ ID No.14、SEQ ID No.16、SEQ IDNo.18、SEQ ID No.20、SEQ ID No.22、SEQ ID No.24氨基酸序列的蛋白質(zhì)的DNA序列，或編碼具有約60％序列同一性并且具有煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中酶活性的蛋白質(zhì)的DNA序列，包括含有選自SEQ ID No.1、SEQ ID No.3、SEQ ID No.5、SEQ ID No.7、SEQ ID No.9、SEQ ID No.11、SEQ ID No.13、SEQ ID No.15、SEQ ID No.17、SEQ IDNo.19、SEQ ID No.21、或SEQ ID No.23核苷酸序列的DNA序列，所述用途為在脅迫條件下增加植物脅迫抗性，或減少活性氧水平，或維持植物或植物細(xì)胞中的NADH水平。
附圖簡述
圖1是面包酵母(釀酒酵母(Saccaromyces cerevisuae))中公知的NAD補(bǔ)救合成途徑和NAD從頭合成途徑的圖示。
圖2-11是多種T-DNA載體的圖示，這些T-DNA載體包含可在植物中表達(dá)的控制元件的控制下編碼NAD補(bǔ)救合成途徑或NAD從頭合成途徑中的酶的DNA區(qū)。使用的縮寫為RBT-DNA右邊界；3’35S來自花椰菜花葉病毒35S轉(zhuǎn)錄物的轉(zhuǎn)錄終止和多聚腺苷酸化作用信號(hào)；Cab22LCab22L轉(zhuǎn)錄物的非翻譯前導(dǎo)序列；P35S2花椰菜花葉病毒35S啟動(dòng)子；3’g7來自根瘤農(nóng)桿菌(Agrobacterium tumefaciens)T-DNA基因7的轉(zhuǎn)錄終止和聚腺甘酸信號(hào)；bar膦絲菌素乙酰轉(zhuǎn)移酶編碼區(qū)；pSSUAra來自擬南芥的核酮糖二磷酸羧化酶加氧酶小亞基轉(zhuǎn)錄物的啟動(dòng)子；LBT-DNA左邊界；Sm/Sp壯觀霉素和鏈霉素抗性基因；pVS1ori適合在農(nóng)桿菌中復(fù)制的VS1起始點(diǎn)；ColE1復(fù)制起點(diǎn)；NLS核定位信號(hào)；PNC1編碼釀酒酵母煙酰胺酶的DNA區(qū)；npt1釀酒酵母煙酸酯磷酸核糖基轉(zhuǎn)移酶；nma1釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶1；nma2釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶2；qns1釀酒酵母NAD合成酶(QNS1)。
發(fā)明詳述本發(fā)明是基于這樣的發(fā)現(xiàn)，即可以使用編碼酵母NAD補(bǔ)救途徑中的植物功能酶的DNA序列獲得轉(zhuǎn)基因植物，所述轉(zhuǎn)基因植物比不包含這些DNA序列的植物對(duì)脅迫特別是對(duì)非生物脅迫的抗性更強(qiáng)。與對(duì)照植物相比，轉(zhuǎn)基因植物在脅迫條件下也表現(xiàn)出活性氧(ROS)水平顯著降低和維持高水平的NADH。
因此在本發(fā)明的一個(gè)實(shí)施方案中，提供了獲得具有增加的脅迫抗性的植物的方法，所述方法包括步驟-將如這里描述的脅迫抗性嵌合基因引入植物細(xì)胞中以獲得含有這種脅迫抗性嵌合基因的細(xì)胞；-再生這些含有脅迫抗性嵌合基因的細(xì)胞以獲得含有脅迫抗性嵌合基因的植物群，和-從這些植物群中選擇植物，所述植物與類似的非轉(zhuǎn)基因植物相比在脅迫條件下表現(xiàn)出增加的脅迫抗性和/或減少的ROS水平和/或維持高水平的NADH。
因此所述脅迫抗性嵌合基因包含有效連接到DNA區(qū)的可在植物中表達(dá)的啟動(dòng)子和參與轉(zhuǎn)錄終止和聚腺甘酸化的3’區(qū)，所述DNA區(qū)編碼煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的植物功能酶，所述酶選自煙酰胺酶、煙酸酯磷酸核糖基轉(zhuǎn)移酶、煙酸單核苷酸腺苷轉(zhuǎn)移酶或煙酰胺腺嘌呤二核苷酸合成酶。
在這里使用“煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的植物功能酶”是指當(dāng)連接到適當(dāng)?shù)目刂圃?如可在植物中表達(dá)的啟動(dòng)子和終止子區(qū))并將其引入到植物中時(shí)，能被轉(zhuǎn)錄和翻譯產(chǎn)生在植物細(xì)胞中有功能的NAD補(bǔ)救合成途徑中的酶。所包括的NAD補(bǔ)救合成途徑中的酶(和編碼基因)不僅從植物來源中獲得，還從酵母(釀酒酵母)或其他酵母或真菌中獲得。認(rèn)為后一種來源的蛋白質(zhì)甚至更適用于本發(fā)明的方法，因?yàn)檫@些蛋白質(zhì)較少可能受到酶反饋調(diào)節(jié)等，而相似的植物來源的酶則容易受酶的反饋調(diào)節(jié)。
參與NAD補(bǔ)救合成途徑的酶包括如下-煙酰胺酶(EC 3.5.1.19)催化煙酰胺上的酰胺基團(tuán)水解，從而釋放出煙酸和NH3。該酶又稱為煙酰胺脫氨酶、煙酰胺酰胺酶、YNDase或煙酰胺酰胺水解酶。
-煙酸酯磷酸核糖基轉(zhuǎn)移酶(EC 2.4.2.11)又稱為煙酸核苷酸酶、煙酸單核苷酸糖苷水解酶、煙酸單核苷酸焦磷酸化酶、煙酸磷酸核糖基轉(zhuǎn)移酶，其催化下面的反應(yīng)煙酸酯-D-核苷酸+二磷酸＝煙酸酯+5-磷酸-α-D-核糖1-二磷酸。
-煙酸酯-核苷酸腺苷轉(zhuǎn)移酶(EC 2.7.7.18)又稱為脫酰胺NAD+焦磷酸化酶、煙酸酯單核苷酸腺苷轉(zhuǎn)移酶、脫酰胺煙酰胺腺嘌呤二核苷酸焦磷酸化酶、NaMT-ATase、煙酸單核苷酸腺苷轉(zhuǎn)移酶，其催化下面的反應(yīng)ATP+煙酸酯核苷酸＝二磷酸+脫酰胺NAD+NAD合酶(EC 6.3.1.5)又稱為NAD合成酶、NAD+合酶、煙酰胺腺嘌呤二核苷酸合成酶、二磷酸吡啶核苷酸合成酶，其催化下面的反應(yīng)脫酰胺NAD++ATP+NH3＝AMP+二磷酸+NAD+在本發(fā)明的一個(gè)實(shí)施方案中，編碼NAD補(bǔ)救合成途徑中不同的酶的編碼區(qū)包含一段核苷酸序列，例如，SEQ ID No 1、SEQ ID No 3、SEQ IDNo 5、SEQ ID No 7或SEQ ID No 9的核苷酸序列，所述核苷酸序列編碼蛋白質(zhì)具有的氨基酸序列如SEQ ID No 2、SEQ ID No 4、SEQ ID No 6、SEQ ID No 8或SEQ ID No 10所列。
但是，應(yīng)該明確的是，也可以使用這些核苷酸序列的變體，包括其插入、缺失和取代，達(dá)到同樣的效果。同樣地，可以使用來自除了釀酒酵母外其他物種的所提到的核苷酸序列的同源物。這些核苷酸序列包括但不限于植物來源的核苷酸序列，編碼具有相同氨基酸序列的蛋白質(zhì)的核苷酸序列，以及這些核苷酸序列的變體。后者的實(shí)例是編碼帶有SEQ ID No 12、14、16、18、20、22或24中列出的氨基酸序列的蛋白質(zhì)的核苷酸序列，例如SEQ ID No 11、13、15、17、19、21或23的核苷酸序列。
所述核苷酸序列的變體與鑒定的編碼NAD補(bǔ)救合成途徑中酶的核苷酸序列(如序列表中鑒定的那些)具有序列同一性，其優(yōu)選至少為約80％，或85％、或90％或95％。優(yōu)選地，這些變體能編碼與NAD補(bǔ)救途徑中的酶具有相同酶活性的有功能的蛋白質(zhì)。為了本發(fā)明的目的，用百分比表示的兩個(gè)相關(guān)核苷酸或氨基酸序列的“序列同一性”指兩個(gè)最佳比對(duì)序列中相同殘基位置的數(shù)目(x100)除以所有比較殘基位置的數(shù)目。空位即比對(duì)中殘基在一個(gè)序列中存在而在另一個(gè)序列中沒有的位置，被認(rèn)為是沒有相同殘基的位置。兩個(gè)序列的比對(duì)用Needleman和Wunsch算法(Needleman和Wunsch 1970)執(zhí)行。上述計(jì)算機(jī)輔助的序列比對(duì)可以用標(biāo)準(zhǔn)軟件程序如GAP方便地執(zhí)行，所述GAP為Wisconsin Package Version 10.1(GeneticsComputer Group，Madision，Wisconsin，USA)的部分，使用帶有空位產(chǎn)生罰分為50和空位延伸罰分為3的缺省得分矩陣。
如Hunt等人所述(見上)，可以通過基因組數(shù)據(jù)芯片(in silico)分析鑒定編碼來自酵母NAD補(bǔ)救途徑的酶的核苷酸序列或編碼來自酵母之外生物體的酶的核苷酸序列的同源核苷酸序列。
同源核苷酸序列也可以使用編碼NAD補(bǔ)救合成途徑中酶的鑒定的核苷酸序列(如在序列表中鑒定的核苷酸序列)作為探針，在嚴(yán)格條件下通過雜交得以鑒定和分離。
如這里所用“嚴(yán)格雜交條件”意指雜交通常發(fā)生在探針和目標(biāo)序列之間的同一性至少為95％，并優(yōu)選至少97％時(shí)。嚴(yán)格雜交條件的實(shí)例是在含有50％甲醛、5×SSC(150mM NaCl、15mM檸檬酸三鈉)、50mM磷酸鈉(pH 7.6)、5×Denhardt雜交溶液、10％硫酸葡聚糖和20μg/ml變性剪切的載體DNA(如鮭魚精子DNA)的溶液中孵育過夜，然后用0.1×SSC在大約65℃洗滌雜交支持物，優(yōu)選洗兩次每次約10分鐘。其他的雜交和洗滌條件是眾所周知的，并且示例于Sambrook等人，Molecular CloningALaboratory Manual，第二版，Cold Spring Harbor，NY(1989)，特別是第11章。
這些變體序列也可以通過DNA擴(kuò)增技術(shù)獲得，其中使用對(duì)于編碼NAD補(bǔ)救途徑中的酶的基因具有特異性的寡核苷酸作為引物，例如但不限于包含約20至約50個(gè)連續(xù)核苷酸的寡核苷酸，所述連續(xù)核苷酸選自SEQID No 1、3、5、7、9、11、13、15、17、19、21、23的核苷酸序列或它們的互補(bǔ)序列。
可以使用本發(fā)明的方法獲得耐受不同種類脅迫誘導(dǎo)條件特別是非生物脅迫條件的植物，所述非生物脅迫條件包括浸沒、高光照條件、高UV輻射水平、增加的過氧化氫水平、干旱條件、高溫或低溫、增加的鹽度條件。本發(fā)明的方法也可以用來降低生長在不利條件下特別是非生物脅迫條件下的植物細(xì)胞中ROS的水平，所述非生物脅迫條件包括浸沒、高光照條件、高UV輻射水平、增加的過氧化氫水平、干旱條件、高溫或低溫、增加的鹽度條件等。可以用在本領(lǐng)域內(nèi)公知的方法(包括在實(shí)施例3中所述的方法)測(cè)定ROS水平或NADH水平。
使用這里所述方法可以獲得植物，其中在非脅迫條件下(例如但不限于低光照條件下)其ROS的水平等于或低于對(duì)照植物。在非脅迫條件下，這些植物中的ROS水平可以是在低光照條件下對(duì)照植物水平的50％至100％，尤其在約60％至約85％之間。在脅迫條件下，這些植物中的ROS水平是脅迫條件下對(duì)照植物的ROS水平的大約50％-80％，相當(dāng)于非脅迫條件下對(duì)照植物中的ROS水平的大約60％-80％。同樣地，在非脅迫條件下，例如但不限于低光照條件下，這些植物的NADH水平等于或高于對(duì)照植物中的水平。在非脅迫條件下，這些植物中的NADH水平是在低光照條件下的對(duì)照植物中NADH水平的100％-160％之間，尤其在大約120％至大約140％之間。在脅迫條件下這些植物中的NADH水平是脅迫條件下對(duì)照植物中NADH水平的大約200％-300％，相當(dāng)于非脅迫條件下對(duì)照植物中ROS水平的大約100％-160％。
獲得轉(zhuǎn)基因植物的方法不是本發(fā)明的關(guān)鍵，任何轉(zhuǎn)化方法和適合具體植物物種的再生方法都可以使用。這些方法是本領(lǐng)域內(nèi)眾所周知的，其包括農(nóng)桿菌介導(dǎo)的轉(zhuǎn)化、基因槍遞送、顯微注射、完整細(xì)胞的電穿孔、聚乙二醇介導(dǎo)的原生質(zhì)體轉(zhuǎn)化、原生質(zhì)體電穿孔、脂質(zhì)體介導(dǎo)的轉(zhuǎn)化、硅須介導(dǎo)的轉(zhuǎn)化(silicon-whiskers mediated transformation)等。然后，用這種方式得到的轉(zhuǎn)化細(xì)胞可以再生為成熟的有繁殖能力的植物。
獲得的轉(zhuǎn)化植物可以用于傳統(tǒng)的育種方案以產(chǎn)生更多的具有相同特征的轉(zhuǎn)化植物，或?qū)⒈景l(fā)明的嵌合基因引入相同或相關(guān)植物物種的其它品種或雜交植物中。從轉(zhuǎn)化植物中得到的種子含有作為穩(wěn)定的基因組插入片段的本發(fā)明的嵌合基因，并且也被包含在本發(fā)明中。
很清楚，這里描述的不同脅迫抗性嵌合基因含有編碼NAD補(bǔ)救途徑中不同酶的DNA區(qū)域，這些嵌合基因可以在一個(gè)植物細(xì)胞或植物中組合以進(jìn)一步提高含有所述嵌合基因的植物的脅迫耐受性。因此，在本發(fā)明的一個(gè)實(shí)施方案中，提供的植物細(xì)胞和植物含有至少兩個(gè)脅迫抗性嵌合基因，每一嵌合基因含有不同的編碼區(qū)。
根據(jù)本發(fā)明的轉(zhuǎn)基因植物細(xì)胞和植物系還可以含有能減少如WO00/04173和PCT/EP2004/003995所述的內(nèi)源PARP和/或PARG基因表達(dá)的嵌合基因?？梢岳缤ㄟ^將本發(fā)明的轉(zhuǎn)基因植物系與含有使PARP和/或PARG基因表達(dá)減少的嵌合基因的轉(zhuǎn)基因植物雜交引入更多的嵌合基因。也可以通過將本發(fā)明的嵌合基因引入或轉(zhuǎn)化到含有使PARP或PARG基因表達(dá)減少的嵌合基因的轉(zhuǎn)基因植物細(xì)胞獲得轉(zhuǎn)基因植物細(xì)胞或植物系，反之亦然。
為了本發(fā)明的目的，啟動(dòng)子是可在植物中表達(dá)的啟動(dòng)子。這里所用術(shù)語“可在植物中表達(dá)的啟動(dòng)子”意指能在植物細(xì)胞中控制(起始)轉(zhuǎn)錄的DNA序列。這包括植物起源的任何啟動(dòng)子，也包括非植物起源的但能在植物細(xì)胞中指導(dǎo)轉(zhuǎn)錄的任何啟動(dòng)子，即病毒或細(xì)菌起源的某些啟動(dòng)子，如CaMV35S(Harpster等人，1988 Mol.Gen.Genet.212，182-190)、地下三葉草病毒啟動(dòng)子No 4或No 7(WO9606932)、或T-DNA基因啟動(dòng)子，還包括組織特異性或器官特異性啟動(dòng)子，其包括但不限于種子特異性啟動(dòng)子(例如WO89/03887)、器官原基特異性啟動(dòng)子(An等人，1996，The Plant Cell 8，15-30)、莖特異性啟動(dòng)子(Keller等人，1988，EMBO J.7，3625-3633)、葉特異性啟動(dòng)子(Hudspeth等人，1989，Plant Mol Biol 12，579-589)、葉肉特異性啟動(dòng)子(如光誘導(dǎo)的核酮糖二磷酸羧化酶加氧酶啟動(dòng)子)、根特異性啟動(dòng)子(Keller等人，1989，Genes Devel.3，1639-1646)、塊莖特異性啟動(dòng)子(Keil等人，1989，EMBO J.8，1323-1330)、維管組織特異性啟動(dòng)子(Peleman等人，1989，Gene 84，359-369)、雄蕊選擇性啟動(dòng)子(WO 89/10396、WO 92/13956)、裂開區(qū)特異性啟動(dòng)子(WO 97/13865)等等。
本發(fā)明的嵌合基因也可以裝備在植物中有功能的核定位信號(hào)(“NLS”)，如SV40 NLS，所述NLS有效地連接到編碼NAD補(bǔ)救途徑中的酶的DNA區(qū)。
讀到這篇文獻(xiàn)，本領(lǐng)域的技術(shù)人員將會(huì)立即認(rèn)識(shí)到，只要獲得其中編碼NAD補(bǔ)救途徑中的酶的內(nèi)源基因更加活躍或以更高水平表達(dá)的植物的天然變體可以獲得增加脅迫抗性的類似效果。這樣的變體植物可以這樣得到對(duì)植物群進(jìn)行誘變，例如但不限于EMS誘變，然后再篩選NAD補(bǔ)救途徑中任何一個(gè)酶或其組合的活性增加的植物。
同樣立即顯而易見的是，可以篩選對(duì)前面提到的一般性脅迫條件(特別是非生物脅迫條件)增加耐受性的很多不同的品種或栽培種，然后將對(duì)脅迫條件增加的耐受性與存在編碼NAD補(bǔ)救途徑中酶的任何內(nèi)源基因的特定等位基因相關(guān)聯(lián)。如果這些物種是性親和的，可以通過雜交將這樣的等位基因引入目的植物中；或者使用這里描述的常規(guī)方法(包括雜交技術(shù)或PCR擴(kuò)增技術(shù))對(duì)其鑒定并用重組DNA技術(shù)引入植物中。用育種技術(shù)引入特別期望的等位基因后可以接著用目的等位基因特異性的分子標(biāo)記檢測(cè)。
認(rèn)為這里描述的方法和手段適合所有植物細(xì)胞和植物，同時(shí)包括雙子葉和單子葉植物細(xì)胞，并且植物包括但不限于棉花、蕓苔屬植物、油料種子油菜、小麥、谷物或玉米、大麥、向日葵、稻、燕麥、甘蔗、大豆、蔬菜(包括菊苣、萵苣、番茄)、煙草、馬鈴薯、甜菜、木瓜、菠蘿、芒果、擬南芥，也包括在園藝、花卉或林業(yè)中使用的植物。
在這里使用的“包含”應(yīng)闡釋為說明陳述特征、整體、步驟或成分的存在，但是并不排除存在或增加一種或多種特征、整體、步驟或成分，或它們組成的組。因此，例如，包含核苷酸或氨基酸序列的核酸或蛋白質(zhì)可以比實(shí)際引述的核酸或蛋白質(zhì)包含更多的核苷酸或氨基酸，即所述實(shí)際引述的核酸或蛋白質(zhì)包含在更大的核酸和蛋白質(zhì)中。含有確定結(jié)構(gòu)或功能的DNA區(qū)的嵌合基因可以包含額外的DNA區(qū)等。
具體實(shí)施例方式
以下非限制性實(shí)施例描述了能增加植物細(xì)胞和植物脅迫抗性的嵌合基因的構(gòu)建和這些基因的使用。
除非在實(shí)施例中另有說明，所有重組DNA技術(shù)都是根據(jù)在Sambrook等人(1989)Molecular CloningA Laboratory Manual，第二版，ColdSpring Harbor Laboratory Press，NY和Ausubel等人(1994)CurrentProtocols in Molecular Biology，Current Protocols，USA第1、2卷中描述的標(biāo)準(zhǔn)方案實(shí)施。植物分子工作的標(biāo)準(zhǔn)材料和方法于R.D.D.Croy編著的由BIOS Scientific Publications Ltd (UK)和Blackwell ScientificPublications，UK聯(lián)合出版的Plant Molecular Biology Labfax(1993)中得到描述。標(biāo)準(zhǔn)分子生物學(xué)技術(shù)的其它參考書包括Sambrook和Russell(2001)Molecular CloningA Laboratory Manual，第三版，Cold Spring HarborLaboratory Press，NY，Brown所著卷I和II(1998)Molecular BiologyLabFax，第二版，Academic Press(UK)。聚合酶鏈?zhǔn)椒磻?yīng)的標(biāo)準(zhǔn)材料和方法見Dieffenbach和Dveksler(1995)PCR PrimerA Laboratory Manual，Cold Spring Harbor Laboratory Press，以及McPherson等人(2000)PCR-BasicsFrom Background to Bench，第一版，Springer Verlag，Germany。
在所有的描述和實(shí)施例中，參考下面的序列SEQ ID No.1釀酒酵母煙酰胺酶(PNC1)的核苷酸序列。
SEQ ID No.2釀酒酵母煙酰胺酶(PNC1)的氨基酸序列。
SEQ ID No.3釀酒酵母煙酸酯磷酸核糖基轉(zhuǎn)移酶(NPT1)的核苷酸序列(互補(bǔ))。
SEQ ID No.4釀酒酵母煙酸酯磷酸核糖基轉(zhuǎn)移酶(NPT1)的氨基酸序列。
SEQ ID No.5釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶1(NMA1)的核苷酸序列。
SEQ ID No.6釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶1(NMA1)的氨基酸序列。
SEQ ID No.7釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶2(NMA2)的核苷酸序列。
SEQ ID No.8釀酒酵母的煙酸單核苷酸腺苷轉(zhuǎn)移酶2(NMA2)的氨基酸序列。
SEQ ID No.9釀酒酵母NAD合成酶(QNS1)的核苷酸序列。
SEQ ID No.10釀酒酵母NAD合成酶(QNS1)的氨基酸序列。
SEQ ID No.11擬南芥煙酰胺酶(同工型1)的核苷酸序列。
SEQ ID No.12擬南芥煙酰胺酶(同工型1)的氨基酸序列。
SEQ ID No.13擬南芥煙酰胺酶(同工型2)的核苷酸序列。
SEQ ID No.14擬南芥煙酰胺酶(同工型2)的氨基酸序列。
SEQ ID No.15擬南芥煙酰胺酶(同工型3)的核苷酸序列。
SEQ ID No.16擬南芥煙酰胺酶(同工型3)的氨基酸序列。
SEQ ID No.17擬南芥煙酸酯磷酸核糖基轉(zhuǎn)移酶(同工型1)的核苷酸序列。
SEQ ID No.18擬南芥煙酸酯磷酸核糖基轉(zhuǎn)移酶(同工型1)的氨基酸序列。
SEQ ID No.19擬南芥煙酸酯磷酸核糖基轉(zhuǎn)移酶(同工型2)的核苷酸序列。
SEQ ID No.20擬南芥煙酸酯磷酸核糖基轉(zhuǎn)移酶(同工型2)的氨基酸序列。
SEQ ID No.21擬南芥煙酸單核苷酸腺苷轉(zhuǎn)移酶的核苷酸序列。
SEQ ID No.22擬南芥煙酸單核苷酸腺苷轉(zhuǎn)移酶的氨基酸序列。
SEQ ID No.23擬南芥NAD合成酶的核苷酸序列。
SEQ ID No.24擬南芥NAD合成酶的氨基酸序列。
SEQ ID No.25T-DNA載體pTVE467的核苷酸序列。
SEQ ID No.26T-DNA載體pTVE468的核苷酸序列。
SEQ ID No.27T-DNA載體pTVE469的核苷酸序列。
SEQ ID No.28T-DNA載體pTVE470的核苷酸序列。
SEQ ID No.29T-DNA載體pTVE496的核苷酸序列。
SEQ ID No.30T-DNA載體pTVE497的核苷酸序列。
SEQ ID No.31T-DNA載體pTVE500的核苷酸序列。
SEQ ID No.32T-DNA載體pTVE501的核苷酸序列。
SEQ ID No.33T-DNA載體pTVE502的核苷酸序列。
SEQ ID No.34T-DNA載體pTVE503的核苷酸序列。
實(shí)施例實(shí)施例1脅迫抗性基因的組裝和引入到植物中。
pTVE467為了增加植物的脅迫抗性，用常規(guī)技術(shù)構(gòu)建的嵌合基因依次含有以下DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)堿基對(duì)的DNA片段；-編碼釀酒酵母煙酰胺酶的DNA片段(SEQ ID NO 1)；-來自花椰菜花葉病毒35S轉(zhuǎn)錄物的3’端非翻譯末端的片段(3’35S)。
可以將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE467(SEQID 25)。T-DNA載體pTVE467如圖2所示。
T-DNA載體pTVE467含有下列分子特征(C)表示互補(bǔ)鏈。

pTVE468構(gòu)建與pTVE467中所示類似的嵌合基因，其中煙酰胺酶裝備了標(biāo)準(zhǔn)核定位信號(hào)。因此，該嵌合基因包含以下有效連接的DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)核苷酸的DNA片段；-編碼含有核定位信號(hào)的肽的約20個(gè)核苷酸的DNA片段；-編碼釀酒酵母煙酰胺酶的DNA片段(SEQ ID NO 1)，由此將核定位信號(hào)融合在閱讀框內(nèi)；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
這一嵌合基因和能提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起被引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE468(SEQ ID26)。T-DNA載體pTVE468如圖3所示。
T-DNA載體pTVE468包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE469為了增加植物的脅迫抗性，用常規(guī)技術(shù)構(gòu)建的嵌合基因依次含有以下DNA片段-來自花椰菜花葉病毒的啟動(dòng)子(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)堿基對(duì)的DNA片段；-編碼釀酒酵母煙酸酯磷酸核糖基轉(zhuǎn)移酶(NPT1；SEQ ID NO 3)的DNA片段；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和能提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE469(SEQ ID27)。T-DNA載體pTVE469如圖4所示。
T-DNA載體pTVE469包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE470構(gòu)建與pTVE469所示類似的嵌合基因，其中酵母煙酸酯磷酸核糖基轉(zhuǎn)移酶裝備了標(biāo)準(zhǔn)核定位信號(hào)。因此，這個(gè)嵌合基因包含以下有效連接的DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)核苷酸的DNA片段；-編碼含有核定位信號(hào)的肽的約20個(gè)核苷酸的DNA片段；-編碼釀酒酵母煙酸酯磷酸核糖基轉(zhuǎn)移酶(NPT1；SEQ ID NO 3)的DNA片段，由此將核定位信號(hào)融合在閱讀框內(nèi)；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE470(SEQ ID28)。T-DNA載體pTVE470如圖5所示。
T-DNA載體pTVE470包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE 496為了增加植物的脅迫抗性，用常規(guī)技術(shù)構(gòu)建成的嵌合基因依次含有以下DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)堿基對(duì)的DNA片段；-編碼釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶(NMA1；SEQ ID NO 5)的DNA片段；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE496(SEQ ID29)。T-DNA載體pTVE496如圖6所示。
T-DNA載體pTVE496包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE497構(gòu)建與pTVE496所示類似的嵌合基因，其中酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶裝備了標(biāo)準(zhǔn)核定位信號(hào)。因此，這個(gè)嵌合基因包含以下有效連接的DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)核苷酸的DNA片段；-編碼含有核定位信號(hào)的肽的約20個(gè)核苷酸的DNA片段；-編碼釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶(NMA1；SEQ ID NO 5)的DNA片段，由此將核定位信號(hào)融合在閱讀框內(nèi)；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE497(SEQ ID30)。T-DNA載體pTVE497如圖7所示。
T-DNA載體pTVE497包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE500為了增加植物的脅迫抗性，用常規(guī)技術(shù)構(gòu)建成的嵌合基因依次含有以下DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)堿基對(duì)的DNA片段；-編碼釀酒酵母中的煙酸單核苷酸腺苷轉(zhuǎn)移酶2(NMA2；SEQ ID NO7)的DNA片段；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE500(SEQ ID31)。T-DNA載體pTVE500如圖8所示。
T-DNA載體pTVE500包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE501構(gòu)建與pTVE500所示類似的嵌合基因，其中釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶2裝備了標(biāo)準(zhǔn)核定位信號(hào)。因此，這個(gè)嵌合基因包含以下有效連接的DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)核苷酸的DNA片段；-編碼含有核定位信號(hào)的肽的約20個(gè)核苷酸的DNA片段；-編碼釀酒酵母煙酸單核苷酸腺苷轉(zhuǎn)移酶2(NMA2；SEQ ID NO 7)的DNA片段，由此將核定位信號(hào)融合在閱讀框內(nèi)；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE501(SEQ ID32)。T-DNA載體pTVE501如圖9所示。
T-DNA載體pTVE501包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE502為了增加植物的脅迫抗性，用常規(guī)技術(shù)構(gòu)建的嵌合基因依次含有以下DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)堿基對(duì)的DNA片段；-編碼釀酒酵母中的NAD合酶(QNS1；SEQ ID NO 9)的DNA片段；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE502(SEQ ID33)。T-DNA載體pTVE502如
圖10所示。
T-DNA載體pTVE502包含下列分子特征(C)表示互補(bǔ)鏈。


pTVE503構(gòu)建與pTVE502所示類似的嵌合基因，其中釀酒酵母中NAD合酶裝備了標(biāo)準(zhǔn)核定位信號(hào)。因此，這個(gè)嵌合基因包含以下有效連接的DNA片段-來自花椰菜花葉病毒的啟動(dòng)子區(qū)域(CaMV35S)；-對(duì)應(yīng)Cab22L非翻譯前導(dǎo)序列的約60個(gè)核苷酸的DNA片段；-編碼含有核定位信號(hào)的肽的約20個(gè)核苷酸的DNA片段；-編碼釀酒酵母NAD合酶(QNS1；SEQ ID NO 9)的DNA片段，由此將核定位信號(hào)融合在閱讀框內(nèi)；-來自花椰菜花葉病毒35S轉(zhuǎn)錄子的3’端非翻譯末端的片段(3’35S)。
將這一嵌合基因和提供對(duì)除草劑膦絲菌素抗性的選擇性標(biāo)記基因一起引入到T-DNA載體的左邊界和右邊界序列之間，產(chǎn)生pTVE503(SEQ ID34)。T-DNA載體pTVE503如
圖11所示。
T-DNA載體pTVE503包含下列分子特征(C)表示互補(bǔ)鏈。

用常規(guī)方法將T-DNA載體引入到含有輔助Ti質(zhì)粒的農(nóng)桿菌菌株中。用如本領(lǐng)域所述農(nóng)桿菌介導(dǎo)的轉(zhuǎn)化方法將嵌合基因引入到擬南芥植物中。
實(shí)施例2對(duì)含有實(shí)施例1所述嵌合基因的轉(zhuǎn)基因擬南芥系的分析如在實(shí)施例1所述獲得的表達(dá)酵母NAD補(bǔ)救途徑基因的轉(zhuǎn)基因擬南芥系(T1代)的種子在含有15mg/L膦絲菌素(PPT)的培養(yǎng)基上萌發(fā)并生長。
擬南芥cv Col-0作為對(duì)照。
所有的植物都用高光照脅迫處理。將在每秒30μEinstein/m2生長的兩周齡的植物轉(zhuǎn)移到每秒250μEinstein/m2(高光照)中處理6小時(shí)；接著在黑暗中8小時(shí)后；再用高光照8小時(shí)。
這種處理后，檢測(cè)所有植物系中的NADH含量和超氧化物自由基的含量，并與在低光照條件下轉(zhuǎn)基因和對(duì)照植物系中相同化合物的測(cè)量相比較。結(jié)果概括于表1。
在高光照條件下轉(zhuǎn)基因植物比對(duì)照植物表現(xiàn)出更高水平的NADH含量，并且在高光照條件下轉(zhuǎn)基因植物比對(duì)照植物產(chǎn)生更少的活性氧。其中在裝備或未裝備核定位信號(hào)的編碼NAD補(bǔ)救途徑酶的構(gòu)建體之間沒有觀察到差異。
還對(duì)轉(zhuǎn)基因植物系對(duì)高光照脅迫條件的耐受性進(jìn)行了表型計(jì)分。為此，將植物在體外低光照條件下(每秒30μEinstein/m2)生長兩周而后轉(zhuǎn)到高光照條件(每秒250μEinstein/m2；16小時(shí)光照-8小時(shí)黑暗)3天。在高光照處理后，將植物轉(zhuǎn)回到低光照條件下，并在表型計(jì)分前再生長3天。
對(duì)照植物矮小，并且已經(jīng)開始開花(脅迫誘導(dǎo))；而含有如實(shí)施例1中所述嵌合基因的轉(zhuǎn)基因植物系則比對(duì)照植物高大并且僅開始抽薹(bolt)。
表1.過表達(dá)如實(shí)施例1所述嵌合酵母基因的轉(zhuǎn)基因擬南芥系的高光照耐受性。

平均值的標(biāo)準(zhǔn)誤差＜10％實(shí)施例3NADH含量和超氧化物含量的檢測(cè)方案用水溶性四氮唑鹽定量細(xì)胞內(nèi)NAD(P)H參考文獻(xiàn)Jun Nakamura，Shoji Asakura，Susan D.Hester，Gilbert de Murcia，Keith W.Caldecott和James A.Swenberg(2003)Quantitation ofintracellular NAD(P)H can monitor an imbalance of DNA single strandbreak repair in base excision repair deficient cells in real time.NucleicAcids Research 31(17)，e104.
植物材料最常用的植物材料-體外生長14-18天但未開花的擬南芥芽-油料種子油菜的下胚軸外植體細(xì)胞計(jì)數(shù)試劑盒-8(CCK-8)Sopachem n.v./比利時(shí)72A，Avenue du Laarbeeklaan-1090 Brussels比利時(shí)內(nèi)含物5mL瓶中含有5mMol/L的WST-8(四氮唑鹽)、0.2mMol/L的1-甲氧基PMS、150mMol/L NaCl反應(yīng)溶液-10ml 25mM磷酸鉀緩沖液pH7.4-0.5ml CCK-8-0.1mM 1-甲氧基-5-甲基吩嗪甲基硫酸鹽(＝1-甲氧基吩嗪硫酸二甲酯)1μL/mL的100mM儲(chǔ)液(分子量＝336.4；2.973ml水中溶100mg)-1滴吐溫20/25ml操作-收集植物材料并置于25mM的磷酸鉀緩沖液(pH7.4)中。
如150個(gè)油料種子油菜的下胚軸外植體1克擬南芥的芽(不帶根)-用反應(yīng)溶液替換緩沖液1克擬南芥苗加15ml
150個(gè)油料種子油菜下胚軸外植體加15ml-在黑暗中于26℃孵育0.5小時(shí)(接著反應(yīng))-在450nm測(cè)量反應(yīng)溶液的吸光度通過定量XTT的減少來測(cè)量超氧化物的產(chǎn)生參考文獻(xiàn)De Block，M.，De Brouwer，D.(2002)A simple and robust in vitro assayto quantify the vigour of oilseed rape lines and hybrids.Plant Physiol.Biochem.40，845-852A.歐洲油菜(BRASSICA NAPUS)培養(yǎng)基和反應(yīng)緩沖液播種培養(yǎng)基(培養(yǎng)基201)半濃縮的Murashige和Skoog鹽2％的蔗糖pH 5.80.6％瓊脂(Difco Bacto瓊脂)250mg/l triacillin愈傷組織誘導(dǎo)培養(yǎng)基A2S3MS培養(yǎng)基、0.5g/l Mes(pH5.8)、3％蔗糖、40mg/l硫酸腺嘌呤、0.5％瓊脂糖、1mg/l 2，4-D、0.25mg/l NAA、1mg/l BAP、250mg/ltriacillin。
反應(yīng)緩沖液25mM磷酸鉀緩沖液pH8，1mM 3’-{1-[苯基氨-羰基]-3，4-四氮唑}-雙(4-甲氧基-6-硝基)鈉＝XTT(BioVectra，Canada)(分子量674.53)通過小心溫?zé)崛芤?±37℃)溶解XTT(用前冷卻到室溫)每25ml緩沖液中加1滴吐溫20
種子消毒-種子預(yù)萌發(fā)-幼苗生長種子在70％的乙醇中浸泡2分鐘，然后用含有0.1％吐溫20的次氯酸鈉溶液(含有約6％的活性氯)表面消毒15分鐘。最后，用1升無菌自來水沖洗種子。將種子在無菌自來水中孵育至少1小時(shí)(讓可能抑制萌發(fā)的成分從種子擴(kuò)散)。
將種子放在含有50毫升無菌自來水(+250mg/l triacillin)的250ml錐形瓶(erlenmeyer flask)中。搖動(dòng)約20小時(shí)。
將長出根的種子從含有約125ml播種培養(yǎng)基的Duchefa(每個(gè)容器10粒種子，不要太多以減少由于污染帶來的種子損失)放到實(shí)驗(yàn)室通風(fēng)容器中。這些種子在±24℃和每天16小時(shí)光照(每秒10-30μEinstein/m2)條件下萌發(fā)。
附為了計(jì)算，必須播種的種子數(shù)目每個(gè)幼苗5個(gè)下胚軸片段下胚軸外植體的預(yù)培養(yǎng)和脅迫的誘導(dǎo)-播種12-14天后，將下胚軸切為約7-10mm片段。
-下胚軸外植體(25個(gè)下胚軸/Optilux培養(yǎng)皿，F(xiàn)alcon S1005，丹麥)在A2S2培養(yǎng)基中在25℃(于每秒10-30μEinstein/m2)條件下培養(yǎng)5天。
附每個(gè)條件使用150個(gè)下胚軸外植體。
-脅迫誘導(dǎo)將下胚軸外植體轉(zhuǎn)移到分別含有0、25和50mg/l乙酰水楊酸的A2S3培養(yǎng)基中。
在25℃和每天16小時(shí)光照(每秒10-30μEinstein/m2)的條件下孵育約24小時(shí)。
XTT測(cè)定-將150個(gè)下胚軸外植體轉(zhuǎn)移到50ml Falcon管中。
-用反應(yīng)緩沖液(無XTT)洗滌。
-加20ml反應(yīng)緩沖液+XTT
(必須浸沒外植體，而不要真空滲入)-在黑暗中于26℃孵育。
-反應(yīng)后，在470nm測(cè)量反應(yīng)培養(yǎng)基的吸光度。
B.擬南芥培養(yǎng)基和反應(yīng)緩沖液植物培養(yǎng)基半濃縮的Murashige和Skoog鹽B5維生素1.5％蔗糖pH 5.80.7％Difco瓊脂孵育培養(yǎng)基半濃縮的MS鹽1％蔗糖0.5g/l MES pH5.8每25ml培養(yǎng)基中加1滴吐溫20反應(yīng)緩沖液25mM磷酸鉀緩沖液pH8.0，1mM 3’-{1-[苯基氨-羰基]-3，4-四氮唑}-二(4-甲氧基-6-硝基)鈉＝XTT(BioVectra，Canada)(分子量674.53)通過小心溫?zé)崛芤?±37℃)溶解XTT(用前冷卻到室溫)每25ml緩沖液中加1滴吐溫20擬南芥植物-擬南芥株系用于測(cè)試的對(duì)照株系(用來獲得測(cè)試株系的母株系)-擬南芥種子的消毒用70％乙醇處理2分鐘用每20ml溶液(含有6％活性氯)中加入1滴吐溫20的溶液漂白10分鐘用無菌自來水沖洗5次附消毒是在2ml的Eppendorf管中進(jìn)行。擬南芥種子沉到管底，使能夠用1ml的自動(dòng)移液器(pipetman)移去液體。
-種子的預(yù)萌發(fā)在直徑為9cm的Optilux培養(yǎng)皿(Flcon)中含有12ml無菌自來水。
低光照過夜到24小時(shí)。
-擬南芥植物的生長將種子播種在含有±125ml植物培養(yǎng)基的Falcon的內(nèi)格(intergrid)組織培養(yǎng)皿(nr.3025)中每格放1粒種子。
植物抽薹前在24℃和16小時(shí)光照(每秒30μEinstein/m2)-8小時(shí)黑暗的條件下生長約18天。
附實(shí)施測(cè)定需要1克的植物材料(不帶根的芽)/株系/條件。1克芽對(duì)應(yīng)于40-60株植物。
脅迫誘導(dǎo)百草枯-收獲擬南芥芽(無根)-將1克芽放在分別含有0、5和10μM百草枯的孵育培養(yǎng)基中(必須浸沒芽，而不要真空滲透)。
孵育培養(yǎng)基在Falcon的內(nèi)格組織培養(yǎng)皿(nr.3025)中±150ml-在24℃黑暗中孵育±24小時(shí)以及每天16小時(shí)光照(每秒30-50μEinstein/m2)。
高光照-將半數(shù)平皿轉(zhuǎn)移到高光照(每秒250μEinstein/m2)并孵育4-20小時(shí)。
XTT測(cè)定-從瓊脂平皿中(高光照脅迫)或從液體孵育培養(yǎng)基中(百草枯脅迫)收獲芽(無根)，并將其放在含反應(yīng)緩沖液(無XTT)的50ml的Falcon管中。
-用含有XTT(15ml/g)的緩沖液替換反應(yīng)緩沖液；-必須浸沒芽，而不要真空滲透；-在黑暗中于26℃孵育；-反應(yīng)后，在470nm處檢測(cè)反應(yīng)培養(yǎng)基的吸光度(約1小時(shí))。
實(shí)施例4過表達(dá)酵母煙酰胺酶(Pnc1)基因增加擬南芥植物對(duì)臭氧的耐受性將嵌合載體pTVE467(實(shí)施例1)用于哥倫比亞生態(tài)型擬南芥的轉(zhuǎn)化。用Southern-DNA和Northern-RNA印跡分析初始轉(zhuǎn)化體。鑒定到一個(gè)轉(zhuǎn)基因系帶有單個(gè)拷貝的Pnc1轉(zhuǎn)基因構(gòu)建體并且具有高穩(wěn)態(tài)水平的轉(zhuǎn)基因全長Pnc1-mRNA(20pg/5μg總RNA)。
萌發(fā)6周之后，將含有單個(gè)拷貝轉(zhuǎn)基因系的植物和作為對(duì)照的哥倫比亞野生型各100株植物在煙熏室內(nèi)暴露于臭氧。在連續(xù)兩天中分別用250、350、500ppb的臭氧濃度處理植物，每天5小時(shí)。處理之后，對(duì)全部植物肉眼篩選表現(xiàn)為壞死斑的臭氧損傷植物。結(jié)果概括于表2。在500ppb臭氧暴露處理中，幾乎所有的植物表現(xiàn)為壞死斑，但在2個(gè)較低臭氧濃度的暴露處理中，轉(zhuǎn)基因植物受傷的百分比顯著減少。
另外，在增加的臭氧濃度下，確定所有轉(zhuǎn)基因株系和野生型植物的生存力表現(xiàn)指數(shù)(PI)的演化。PI可以通過公式計(jì)算PI＝(ABS/CS)×(TR/CS)×(ET/CS)(ABS＝由天線色素Chi*吸收的光子流量；CS＝橫斷面；TR＝由反應(yīng)中心捕獲的并轉(zhuǎn)化為氧化還原能的能量；ET＝進(jìn)一步流向下游引起CO2固定的電子流量)。轉(zhuǎn)基因株系的生存力表現(xiàn)指數(shù)PI隨著臭氧濃度的增加顯著地增加，而野生型植物的生存力表現(xiàn)指數(shù)隨著臭氧濃度的增加保持恒定。這可以解釋為在轉(zhuǎn)基因株系中為對(duì)抗臭氧傷害產(chǎn)生的補(bǔ)償性生理應(yīng)答。
表2過表達(dá)酵母煙酰胺酶(Pnc1)基因的擬南芥植物對(duì)臭氧的耐受性增加

*呈現(xiàn)壞死斑的植物的百分比此外，還將對(duì)照植物，含有嵌合基因Pnc1的純合子轉(zhuǎn)基因植物種群以及雜合子轉(zhuǎn)基因種群用臭氧煙熏處理，并計(jì)錄了肉眼可見的傷害以及與未煙熏植物相比的多種生理應(yīng)答。所述評(píng)估包括非調(diào)節(jié)熒光、調(diào)節(jié)熒光、葉綠素的測(cè)量以及新增重量的確定。
根據(jù)肉眼可見的傷害和生理應(yīng)答，對(duì)每個(gè)種群定級(jí)表明臭氧影響的程度。評(píng)估越是負(fù)值，種群對(duì)臭氧的應(yīng)答越敏感。
對(duì)照非轉(zhuǎn)基因種群和雜合子轉(zhuǎn)基因種群的累計(jì)得分為-13，而兩個(gè)純合子轉(zhuǎn)基因種群的得分分別是-6和-2。因此，很明顯，純合子轉(zhuǎn)基因種群在統(tǒng)計(jì)學(xué)上的表現(xiàn)顯著優(yōu)于對(duì)照植物。
序列表<110>拜爾生物科學(xué)公司<120>脅迫抗性植物<130>BCS04-2005 WO1<150>EP 04077624.7<151>2004-09-24<150>US 60/628826<151>2004-11-17<160>34<170>PatentIn version 3.0<210>1<211>651<212>DNA<213>釀酒酵母(Saccharomyces cerevisiae)<400>1atgaagactt taattgttgt tgatatgcaa aatgatttta tttcaccttt aggttccttg 60actgttccaa aaggtgagga attaatcaat cctatctcgg atttgatgca agatgctgat120agagactggc acaggattgt ggtcaccaga gattggcacc cttccagaca tatttcgttc180gcaaagaacc ataaagataa agaaccctat tcaacataca cctaccactc tccaaggcca240ggcgatgatt ccacgcaaga gggtattttg tggcccgtac actgtgtgaa aaacacctgg300ggtagtcaat tggttgacca aataatggac caagtggtca ctaagcatat taagattgtc360gacaagggtt tcttgactga ccgtgaatac tactccgcct tccacgacat ctggaacttc420cataagaccg acatgaacaa gtacttagaa aagcatcata cagacgaggt ttacattgtc480ggtgtagctt tggagtattg tgtcaaagcc accgccattt ccgctgcaga actaggttat540aagaccactg tcctgctgga ttacacaaga cccatcagcg atgatcccga agtcatcaat600aaggttaagg aagagttgaa ggcccacaac atcaatgtcg tggataaata a 651<210>2<211>216<212>PRT<213>釀酒酵母<400>2Met Lys Thr Leu Ile Val Val Asp Met Gln Asn Asp Phe Ile Ser Pro1 5 10 15Leu Gly Ser Leu Thr Val Pro Lys Gly Glu Glu Leu Ile Asn Pro Ile20 25 30Ser Asp Leu Met Gln Asp Ala Asp Arg Asp Trp His Arg Ile Val Val35 40 45Thr Arg Asp Trp His Pro Ser Arg His Ile Ser Phe Ala Lys Asn His50 55 60
Lys Asp Lys Glu Pro Tyr Ser Thr Tyr Thr Tyr His Set Pro Arg Pro65 70 75 80Gly Asp Asp Ser Thr Gln Glu Gly Ile Leu Trp Pro Val His Cys Val85 90 95Lys Asn Thr Trp Gly Ser Gln Leu Val Asp Gln Ile Met Asp Gln Val100 105 110Val Thr Lys His Ile Lys Ile Val Asp Lys Gly Phe Leu Thr Asp Arg115 120 125Glu Tyr Tyr Ser Ala Phe His Asp Ile Trp Asn Phe His Lys Thr Asp130 135 140Met Asn Lys Tyr Leu Glu Lys His His Thr Asp Glu Val Tyr Ile Val145 150 155 160Gly Val Ala Leu Glu Tyr Cys Val Lys Ala Thr Ala Ile Ser Ala Ala165 170 175Glu Leu Gly Tyr Lys Thr Thr Val Leu Leu Asp Tyr Thr Arg Pro Ile180 185 190Ser Asp Asp Pro Glu Val Ile Asn Lys Val Lys Glu Glu Leu Lys Ala195 200 205His Asn Ile Asn Val Val Asp Lys210 215<210>3<211>1290<212>DNA<213>釀酒酵母<400>3ttaggtccat ctgtgcgctt cgttatcacc actccaactt cgttcagtat atcccaattc 60ctctttcact ctcttcacag tggcaggatc tcccatattt ttacctaagt tatcagaaat 120tttgatagcg tgattaccat ttacttctaa tagtttgata acgatgttta acggctcact 180tttaacctgg ggttctgact tcttacgaaa atcattagta aagtttgtgc caataccgaa 240tgtggctagc attccattct ctttagctgc atgggagtaa gttattgcct tttcgacgtt 300caaagaatcg gaataacaga taatcttcga gaatttaggc aatttcaaca cgtcatggta 360atggtgggaa atctttttgg tatactcaac tgggtctcca gaatcttgtc taacaccgac 420gtaagcatca gaatatggtg gacggaatga ttttaaaaag tcatcagttc caaaagtatc 480cgttaatgct aaaccagcat tttttgcacc aaaagtattg atccaacaat ccattgcatt 540tttattggca tgcaaataat cttcactaat agaagcgact cccataaccc actcgtgagc 600cacagtaccg attggcttga ctccatattt cttggcaaat aaaatatttg atgtgcctaa 660taatagcgat ttgtttctgt ctgggttacc gttcacagct ttcatgattc cttgcataat 720tagatcttga gccttcagag atctacgacg tcttgtacca aattcactga atctaatacc 780attatcaaac aaagtttccg ccttcttctc agcttgttct aattggtttt cgtagtccca 840gtcgatgtca acaaatttaa aatacgcttc tgatattagg gacagtaagg ggatctcata 900aaggatagta tccttccaac taccactgac taaaattttc aatttgtagt gggtgggctt 960gccctcgatt tcttctgaag tgaaggaaat ctgctcttca gggtgtagtt tgtaattaga1020actgctaata tacttaatat atgccgatgg caaatatggg atttcctgtt ttaagtattc1080aatttcctct tctgtgaacc tcaaatttcc caaatacgaa aattgctctt tcaaccaatt1140aatggcttcc ttattgaagg tcaattggga cgacctgttg gtatatttat aagtaactgt1200aacatctgga aaattagtga agacagcagc atgcatcgta atcttgtaca tgtctgtgtc1260caaaagagac tttatcactg gttctgacat 1290
<210>4<211>429<212>PRT<213>釀酒酵母<400>4Met Ser Glu Pro Val Ile Lys Ser Leu Leu Asp Thr Asp Met Tyr Lys1 5 10 15Ile Thr Met His Ala Ala Val Phe Thr Asn Phe Pro Asp Val Thr Val20 25 30Thr Tyr Lys Tyr Thr Asn Arg Ser Ser Gln Leu Thr Phe Asn Lys Glu35 40 45Ala Ile Asn Trp Leu Lys Glu Gln Phe Ser Tyr Leu Gly Asn Leu Arg50 55 60Phe Thr Glu Glu Glu Ile Glu Tyr Leu Lys Gln Glu Ile Pro Tyr Leu65 70 75 80Pro Ser Ala Tyr Ile Lys Tyr Ile Ser Ser Ser Asn Tyr Lys Leu His85 90 95Pro Glu Glu Gln Ile Ser Phe Thr Ser Glu Glu Ile Glu Gly Lys Pro100 105 110Thr His Tyr Lys Leu Lys Ile Leu Val Ser Gly Ser Trp Lys Asp Thr115 120 125Ile Leu Tyr Glu Ile Pro Leu Leu Ser Leu Ile Ser Glu Ala Tyr Phe130 135 140Lys Phe Val Asp Ile Asp Trp Asp Tyr Glu Asn Gln Leu Glu Gln Ala145 150 155 160Glu Lys Lys Ala Glu Thr Leu Phe Asp Asn Gly Ile Arg Phe Ser Glu165 170 175Phe Gly Thr Arg Arg Arg Arg Ser Leu Lys Ala Gln Asp Leu Ile Met180 185 190Gln Gly Ile Met Lys Ala Val Asn Gly Asn Pro Asp Arg Asn Lys Ser195 200 205Leu Leu Leu Gly Thr Ser Asn Ile Leu Phe Ala Lys Lys Tyr Gly Val210 215 220Lys Pro Ile Gly Thr Val Ala His Glu Trp Val Met Gly Val Ala Ser225 230 235 240Ile Ser Glu Asp Tyr Leu His Ala Asn Lys Asn Ala Met Asp Cys Trp245 250 255Ile Asn Thr Phe Gly Ala Lys Asn Ala Gly Leu Ala Leu Thr Asp Thr260 265 270Phe Gly Thr Asp Asp Phe Leu Lys Ser Phe Arg Pro Pro Tyr Ser Asp275 280 285Ala Tyr Val Gly Val Arg Gln Asp Ser Gly Asp Pro Val Glu Tyr Thr290 295 300Lys Lys Ile Ser His His Tyr His Asp Val Leu Lys Leu Pro Lys Phe305 310 315 320Ser Lys Ile Ile Cys Tyr Ser Asp Ser Leu Asn Val Glu Lys Ala Ile325 330 335Thr Tyr Ser His Ala Ala Lys Glu Asn Gly Met Leu Ala Thr Phe Gly340 345 350Ile Gly Thr Asn Phe Thr Asn Asp Phe Arg Lys Lys Ser Glu Pro Gln355 360 365
Val Lys Ser Glu Pro Leu Asn Ile Val Ile Lys Leu Leu Glu Val Asn370 375 380Gly Asn His Ala Ile Lys Ile Ser Asp Asn Leu Gly Lys Asn Met Gly385 390 395 400Asp Pro Ala Thr Val Lys Arg Val Lys Glu Glu Leu Gly Tyr Thr Glu405 410 415Arg Ser Trp Ser Gly Asp Asn Glu Ala His Arg Trp Thr420 425<210>5<211>1206<212>DNA<213>釀酒酵母<400>5atggatccca caagagctcc ggatttcaaa ccgccatctg cagacgagga attgattcct 60ccacccgacc cggaatctaa aattcccaaa tctattccaa ttattccata cgtcttagcc120gatgcgaatt cctctataga tgcacctttt aatattaaga ggaagaaaaa gcatcctaag180catcatcatc accatcatca cagtcgtaaa gaaggcaatg ataaaaaaca tcagcatatt240ccattgaacc aagacgactt tcaaccactt tccgcagaag tgtcttccga agatgatgac300gcggatttta gatccaagga gagatacggt tcagattcaa ccacagaatc agaaactaga360ggtgttcaga aatatcagat tgctgattta gaagaagttc cacatggaat cgttcgtcaa420gcaagaacct tggaagacta cgaattcccc tcacacagat tatcgaaaaa attactggat480ccaaataaac tgccgttagt aatagtagca tgtgggtctt tttcaccaat cacctacttg540catctaagaa tgtttgaaat ggctttagat gcaatctctg aacaaacaag gtttgaagtc600ataggtggat attactcccc tgttagtgat aactatcaaa agcaaggctt ggccccatcc660taccatagag tacgtatgtg tgaattggcc tgcgaaagaa cctcatcttg gttgatggtg720gatgcatggg agtcattgca accttcatac acaagaactg ccaaggtctt ggatcatttc780aatcacgaaa tcaatattaa gagaggtggt gtagctactg ttactggaga aaaaattggt840gtgaaaataa tgttgctggc tggtggtgac ctaatagagt caatgggtga accaaacgtt900tgggcggacg ccgatttaca tcacattctc ggtaattacg gttgtttgat tgtcgaacgt960actggttctg atgtaaggtc ttttttgtta tcccatgata ttatgtatga acatagaagg 1020aatattctta tcatcaagca actcatctat aatgatattt cttccacgaa agttcgtcta 1080tttatcagac gcgccatgtc tgtacaatat ttgttaccta attcggtcat caggtatatc 1140caagaacata gactatatgt ggaccaaacc gaacctgtta agcaagttct tggaaacaaa 1200gaatga 1206<210>6<211>401<212>PRT<213>釀酒酵母<400>6Met Asp Pro Thr Arg Ala Pro Asp Phe Lys Pro Pro Ser Ala Asp Glu1 5 10 15Glu Leu Ile Pro Pro Pro Asp Pro Glu Ser Lys Ile Pro Lys Ser Ile20 25 30Pro Ile Ile Pro Tyr Val Leu Ala Asp Ala Asn Ser Ser Ile Asp Ala35 40 45
Pro Phe Asn Ile Lys Arg Lys Lys Lys His Pro Lys His His His His50 55 60His His His Ser Arg Lys Glu Gly Asn Asp Lys Lys His Gln His Ile65 70 75 80Pro Leu Asn Gln Asp Asp Phe Gln Pro Leu Ser Ala Glu Val Ser Ser85 90 95Glu Asp Asp Asp Ala Asp Phe Arg Ser Lys Glu Arg Tyr Gly Ser Asp100 105 110Ser Thr Thr Glu Ser Glu Thr Arg Gly Val Gln Lys Tyr Gln Ile Ala115 120 125Asp Leu Glu Glu Val Pro His Gly Ile Val Arg Gln Ala Arg Thr Leu130 135 140Glu Asp Tyr Glu Phe Pro Ser His Arg Leu Ser Lys Lys Leu Leu Asp145 150 155 160Pro Asn Lys Leu Pro Leu Val Ile Val Ala Cys Gly Ser Phe Ser Pro165 170 175Ile Thr Tyr Leu His Leu Arg Met Phe Glu Met Ala Leu Asp Ala Ile180 185 190Ser Glu Gln Thr Arg Phe Glu Val Ile Gly Gly Tyr Tyr Ser Pro Val195 200 205Ser Asp Asn Tyr Gln Lys Gln Gly Leu Ala Pro Ser Tyr His Arg Val210 215 220Arg Met Cys Glu Leu Ala Cys Glu Arg Thr Ser Ser Trp Leu Met Val225 230 235 240Asp Ala Trp Glu Ser Leu Gln Pro Ser Tyr Thr Arg Thr Ala Lys Val245 250 255Leu Asp His Phe Asn His Glu Ile Asn Ile Lys Arg Gly Gly Val Ala260 265 270Thr Val Thr Gly Glu Lys Ile Gly Val Lys Ile Met Leu Leu Ala Gly275 280 285Gly Asp Leu Ile Glu Ser Met Gly Glu Pro Asn Val Trp Ala Asp Ala290 295 300Asp Leu His His Ile Leu Gly Asn Tyr Gly Cys Leu Ile Val Glu Arg305 310 315 320Thr Gly Ser Asp Val Arg Ser Phe Leu Leu Ser His Asp Ile Met Tyr325 330 335Glu His Arg Arg Asn Ile Leu Ile Ile Lys Gln Leu Ile Tyr Asn Asp340 345 350Ile Ser Ser Thr Lys Val Arg Leu Phe Ile Arg Arg Ala Met Ser Val355 360 365Gln Tyr Leu Leu Pro Asn Ser Val Ile Arg Tyr Ile Gln Glu His Arg370 375 380Leu Tyr Val Asp Gln Thr Glu Pro Val Lys Gln Val Leu Gly Asn Lys385 390 395 400Glu<210>7<211>1188<212>DNA<213>釀酒酵母
<400>7atggatccca ccaaagcacc cgattttaaa ccgccacagc caaatgaaga actacaacca 60ccgccagatc caacacatac gataccaaaa tctggaccca tagttccata tgttttagct120gattataatt cttcgatcga tgctcctttc aatctcgaca tttacaaaac cctgtcgtca180aggaaaaaaa acgccaactc aagcaaccga atggaccata ttccattaaa tactagtgac240ttccagccac tatctcggga tgtatcatcg gaggaggaaa gtgaagggca atcgaatgga300attgacgcta ctctacagga tgttacgatg actgggaatt tgggggtact gaagagccaa360attgctgatt tggaagaagt tcctcacaca attgtaagac aagccagaac tattgaagat420tacgaatttc ctgtacacag attgacgaaa aagttacaag atcctgaaaa actgcctctg480atcatcgttg cttgtggatc attttctccc ataacatacc tacatttgag aatgtttgaa540atggctttag atgatatcaa tgagcaaacg cgttttgaag tggttggtgg ttatttttct600ccagtaagtg ataactatca aaagcgaggg ttagccccag cttatcatcg tgtccgcatg660tgcgaattag catgcgagcg gacatcatct tggttaatgg ttgatgcctg ggaatcttta720caatcaagtt atacaaggac agcaaaagtc ttggaccatt tcaatcatga aataaatatc780aagagaggtg gaatcatgac tgtagatggt gaaaaaatgg gcgtaaaaat catgttattg840gcaggcggtg atcttatcga atccatgggc gagcctcatg tgtgggctga ttcagacctg900caccatattt tgggtaatta tggatgtttg atcgtggaaa ggactggttc tgatgttagg960tccttcttgc tttcccatga tatcatgtat gaacacagaa gaaatatcct tattatcaaa 1020caacttattt acaatgatat ttcctctacg aaagtgcggc ttttcatcag acgtggaatg 1080tcagttcaat atcttcttcc aaactctgtc atccgttaca tccaagagta taatctatac 1140attaatcaaa gtgaaccggt caagcaggtc ttggatagca aagagtga1188<210>8<211>395<212>PRT<213>釀酒酵母<400>8Met Asp Pro Thr Lys Ala Pro Asp Phe Lys Pro Pro Gln Pro Asn Glu1 5 10 15Glu Leu Gln Pro Pro Pro Asp Pro Thr His Thr Ile Pro Lys Ser Gly20 25 30Pro Ile Val Pro Tyr Val Leu Ala Asp Tyr Asn Ser Ser Ile Asp Ala35 40 45Pro Phe Asn Leu Asp Ile Tyr Lys Thr Leu Ser Ser Arg Lys Lys Asn50 55 60Ala Asn Ser Ser Asn Arg Met Asp His Ile Pro Leu Asn Thr Ser Asp65 70 75 80Phe Gln Pro Leu Ser Arg Asp Val Ser Ser Glu Glu Glu Ser Glu Gly85 90 95Gln Ser Asn Gly Ile Asp Ala Thr Leu Gln Asp Val Thr Met Thr Gly100 105 110Asn Leu Gly Val Leu Lys Ser Gln Ile Ala Asp Leu Glu Glu Val Pro115 120 125His Thr Ile Val Arg Gln Ala Arg Thr Ile Glu Asp Tyr Glu Phe Pro130 135 140Val His Arg Leu Thr Lys Lys Leu Gln Asp Pro Glu Lys Leu Pro Leu145 150 155 160Ile Ile Val Ala Cys Gly Ser Phe Ser Pro Ile Thr Tyr Leu His Leu165 170 175
Arg Met Phe Glu Met Ala Leu Asp Asp Ile Asn Glu Gln Thr Arg Phe180 185 190Glu Val Val Gly Gly Tyr Phe Ser Pro Val Ser Asp Asn Tyr Gln Lys195 200 205Arg Gly Leu Ala Pro Ala Tyr His Arg Val Arg Met Cys Glu Leu Ala210 215 220Cys Glu Arg Thr Ser Ser Trp Leu Met Val Asp Ala Trp Glu Ser Leu225 230 235 240Gln Ser Ser Tyr Thr Arg Thr Ala Lys Val Leu Asp His Phe Asn His245 250 255Glu Ile Asn Ile Lys Arg Gly Gly Ile Met Thr Val Asp Gly Glu Lys260 265 270Met Gly Val Lys Ile Met Leu Leu Ala Gly Gly Asp Leu Ile Glu Ser275 280 285Met Gly Glu Pro His Val Trp Ala Asp Ser Asp Leu His His Ile Leu290 295 300Gly Asn Tyr Gly Cys Leu Ile Val Glu Arg Thr Gly Ser Asp Val Arg305 310 315 320Ser Phe Leu Leu Ser His Asp Ile Met Tyr Glu His Arg Arg Asn Ile325 330 335Leu Ile Ile Lys Gln Leu Ile Tyr Asn Asp Ile Ser Ser Thr Lys Val340 345 350Arg Leu Phe Ile Arg Arg Gly Met Ser Val Gln Tyr Leu Leu Pro Asn355 360 365Ser Val Ile Arg Tyr Ile Gln Glu Tyr Asn Leu Tyr Ile Asn Gln Ser370 375 380Glu Pro Val Lys Gln Val Leu Asp Ser Lys Glu385 390 395<210>9<211>2145<212>DNA<213>釀酒酵母<400>9atgtcacatc ttatcacttt agctacatgc aacttgaatc aatgggccct agattttgaa 60ggtaatagag accgtatcct acagtccatt aagattgcca aagagagggg tgccaggtta120cgtgtcggcc cagaactgga aataactggc tacggatgtt tagatcattt tttagaaaat180gacgtttgcc ttcattcatg ggaaatgtat gctcaaatca ttaagaataa agaaacccat240ggattaatac ttgacattgg tatgcccgtt ctacacaaga atgttcgtta taattgtcgt300ttgttatcct tggatggtga gatattgttc ataagaccta agatttggtt agctaatgat360ggtaactata gggaaatgag atttttcaca ccttggatga aacctggcgt ggtggaggac420tttatccttc cacctgagat tcagaaagtt accggccaga gacttgtgcc atttggggac480gctgtgataa attcattgga tacatgcatt ggtacagaaa cttgtgaaga attgtttaca540cctcaatccc cccacatcgc catgtcttta gatggtgtgg aaatcatgac aaactcatct600ggttctcatc atgaactgcg taagttaaat aaaaggttag acctaatttt aaatgccact660aaacgttgtg gtggtgttta cttgtatgca aatcaaagag gttgtgatgg tgacagatta720tattatgatg gctgtgcact aattgccatc aatggtacaa ttgtagccca aggttcacaa780ttttcgctag atgatgtgga agtagttact gctactgtgg acctagaaga ggtgaggagt840tatcgtgcag ctgtcatgtc tcgtggccta caagcctcct tggcagaaat aaagttcaag900cgtattgata ttcctgtaga attggcttta atgacctcca gatttgatcc tacagtgtgt960
ccaacaaaag tccgcgagcc tttctatcac tctcctgagg aagaaattgc actgggacct1020gcttgctgga tgtgggatta tttaagacgt tgtaacggaa cagggttttt ccttccctta1080tctgggggca ttgactcttg tgcaactgca atgattgtcc actctatgtg ccgtttagtg1140accgacgctg ctcaaaatgg aaatgagcaa gttatcaaag acgttcgtaa gataacacgt1200agcggcgatg attggattcc agacagtcca caggatctag cctcaaaaat atttcactcc1260tgtttcatgg gtacggaaaa ttcatccaag gagacaagaa acagagcaaa ggacctttcc1320aatgcaattg gatcttacca cgtggattta aagatggact cattggtatc cagtgtggtg1380tccttattcg aagtagccac tggcaaaaaa ccaatataca aaatatttgg gggatctcaa1440atcgagaact tggctttaca aaacatccag gcgcgtctaa gaatggttct ttcttatctt1500tttgcgcaac tgttgccgtg ggttcgtggt atcccaaact cgggtggatt gttagtactt1560ggtagcgcaa atgttgatga gtgcttacgt gggtatctaa caaaatatga ctgctcctcc1620gcagatatca accctattgg gggtatttca aaaactgact tgaaaagatt cattgcctac1680gcatcaaaac aatataacat gccaatcttg aatgactttt taaacgctac accaactgca1740gaattagaac ctatgactaa agattacgtt caatcggatg agatagatat ggggatgacg1800tatgaagaat tgggcgtgtt tggttaccta agaaaggttg aaaaatgtgg tccttattct1860atgttcttaa aacttcttca tcaatggtcc ccaaagttaa cacctcgtca aatatctgaa1920aaggtgaaaa gatttttctt cttctatgcc atcaacagac acaagcaaac tgttttaact1980cctagttatc atgctgaaca gtattcacca gaagacaaca gatttgactt acgtcctttc2040ttaatcaacc caagatttcc atgggcttca agaaaaattg atgaagttgt cgagcagtgt2100gaagcacata aaggctcaac gcttgacatt atgtctattg attag2145<210>10<211>714<212>PRT<213>釀酒酵母<400>10Met Ser His Leu Ile Thr Leu Ala Thr Cys Asn Leu Asn Gln Trp Ala1 5 10 15Leu Asp Phe Glu Gly Asn Arg Asp Arg Ile Leu Gln Ser Ile Lys Ile20 25 30Ala Lys Glu Arg Gly Ala Arg Leu Arg Val Gly Pro Glu Leu Glu Ile35 40 45Thr Gly Tyr Gly Cys Leu Asp His Phe Leu Glu Asn Asp Val Cys Leu50 55 60His Ser Trp Glu Met Tyr Ala Gln Ile Ile Lys Asn Lys Glu Thr His65 70 75 80Gly Leu Ile Leu Asp Ile Gly Met Pro Val Leu His Lys Asn Val Arg85 90 95Tyr Asn Cys Arg Leu Leu Ser Leu Asp Gly Glu Ile Leu Phe Ile Arg100 105 110Pro Lys Ile Trp Leu Ala Asn Asp Gly Asn Tyr Arg Glu Met Arg Phe115 120 125Phe Thr Pro Trp Met Lys Pro Gly Val Val Glu Asp Phe Ile Leu Pro130 135 140Pro Glu Ile Gln Lys Val Thr Gly Gln Arg Leu Val Pro Phe Gly Asp145 150 155 160Ala Val Ile Asn Ser Leu Asp Thr Cys Ile Gly Thr Glu Thr Cys Glu165 170 175Glu Leu Phe Thr Pro Gln Ser Pro His Ile Ala Met Ser Leu Asp Gly180 185 190
Val Glu Ile Met Thr Asn Ser Ser Gly Ser His His Glu Leu Arg Lys195 200 205Leu Asn Lys Arg Leu Asp Leu Ile Leu Asn Ala Thr Lys Arg Cys Gly210 215 220Gly Val Tyr Leu Tyr Ala Asn Gln Arg Gly Cys Asp Gly Asp Arg Leu225 230 235 240Tyr Tyr Asp Gly Cys Ala Leu Ile Ala Ile Asn Gly Thr Ile Val Ala245 250 255Gln Gly Ser Gln Phe Ser Leu Asp Asp Val Glu Val Val Thr Ala Thr260 265 270Val Asp Leu Glu Glu Val Arg Ser Tyr Arg Ala Ala Val Met Ser Arg275 280 285Gly Leu Gln Ala Ser Leu Ala Glu Ile Lys Phe Lys Arg Ile Asp Ile290 295 300Pro Val Glu Leu Ala Leu Met Thr Ser Arg Phe Asp Pro Thr Val Cys305 310 315 320Pro Thr Lys Val Arg Glu Pro Phe Tyr His Ser Pro Glu Glu Glu Ile325 330 335Ala Leu Gly Pro Ala Cys Trp Met Trp Asp Tyr Leu Arg Arg Cys Asn340 345 350Gly Thr Gly Phe Phe Leu Pro Leu Ser Gly Gly Ile Asp Ser Cys Ala355 360 365Thr Ala Met Ile Val His Ser Met Cys Arg Leu Val Thr Asp Ala Ala370 375 380Gln Asn Gly Asn Glu Gln Val Ile Lys Asp Val Arg Lys Ile Thr Arg385 390 395 400Ser Gly Asp Asp Trp Ile Pro Asp Ser Pro Gln Asp Leu Ala Ser Lys405 410 415Ile Phe His Ser Cys Phe Met Gly Thr Glu Asn Ser Ser Lys Glu Thr420 425 430Arg Asn Arg Ala Lys Asp Leu Ser Asn Ala Ile Gly Ser Tyr His Val435 440 445Asp Leu Lys Met Asp Ser Leu Val Ser Ser Val Val Ser Leu Phe Glu450 455 460Val Ala Thr Gly Lys Lys Pro Ile Tyr Lys Ile Phe Gly Gly Ser Gln465 470 475 480Ile Glu Asn Leu Ala Leu Gln Asn Ile Gln Ala Arg Leu Arg Met Val485 490 495Leu Ser Tyr Leu Phe Ala Gln Leu Leu Pro Trp Val Arg Gly Ile Pro500 505 510Asn Ser Gly Gly Leu Leu Val Leu Gly Ser Ala Asn Val Asp Glu Cys515 520 525Leu Arg Gly Tyr Leu Thr Lys Tyr Asp Cys Ser Ser Ala Asp Ile Asn530 535 540Pro Ile Gly Gly Ile Ser Lys Thr Asp Leu Lys Arg Phe Ile Ala Tyr545 550 555 560Ala Ser Lys Gln Tyr Asn Met Pro Ile Leu Asn Asp Phe Leu Asn Ala565 570 575Thr Pro Thr Ala Glu Leu Glu Pro Met Thr Lys Asp Tyr Val Gln Ser580 585 590
Asp Glu Ile Asp Met Gly Met Thr Tyr Glu Glu Leu Gly Val Phe Gly595 600 605Tyr Leu Arg Lys Val Glu Lys Cys Gly Pro Tyr Ser Met Phe Leu Lys610 615 620Leu Leu His Gln Trp Ser Pro Lys Leu Thr Pro Arg Gln Ile Ser Glu625 630 635 640Lys Val Lys Arg Phe Phe Phe Phe Tyr Ala Ile Asn Arg His Lys Gln645 650 655Thr Val Leu Thr Pro Ser Tyr His Ala Glu Gln Tyr Ser Pro Glu Asp660 665 670Asn Arg Phe Asp Leu Arg Pro Phe Leu Ile Asn Pro Arg Phe Pro Trp675 680 685Ala Ser Arg Lys Ile Asp Glu Val Val Glu Gln Cys Glu Ala His Lys690 695 700Gly Ser Thr Leu Asp Ile Met Ser Ile Asp705 710<210>11<211>597<212>DNA<213>擬南芥(Arabidopsis thaliana)<400>11atggcttcct catcaacgag aaagtacgag acacgaaagc gagatccaaa ctctaaaatc 60gcagctcttc tcgttatcga catgcagaat cacttctcct ccatggccaa acccatcctc120aacaacgttc tcaccaccat cgacatctgc cgacgcgcct cagtccccgt attctttacg180cgtcacaacc acaaatcccc gaccgaccac ggcatgctcg gcgagtggtg taacggcgat240gtaatccttg acggaaccac cgattctgaa atcatccagg agatacaagg ccaagtaacc300ggaccagacg agatggtgga gaagaacacg tacagtgcgt ttaacaaaac ccgcctccag360gaaaacctgg aaaagatcgg agtaaaggag gtgatcgtga tcggagtgat gacgaacttg420tgctgtgaga caacggcgcg tgaagcgttt attaagggtt ttagggtttt tttctcgacg480gacgcgactg cgacgtttaa tgaggagctt cacgaggcta cgctaatgaa tctcgctttt540ggcttcgctt atctcgtcga ttgcgataaa ctccggcgaa gtctactcgg taactaa 597<210>12<211>198<212>PRT<213>擬南芥<400>12Met Ala Ser Ser Ser Thr Arg Lys Tyr Glu Thr Arg Lys Arg Asp Pro1 5 10 15Asn Ser Lys Ile Ala Ala Leu Leu Val Ile Asp Met Gln Asn His Phe20 25 30Ser Ser Met Ala Lys Pro Ile Leu Asn Asn Val Leu Thr Thr Ile Asp35 40 45Ile Cys Arg Arg Ala Ser Val Pro Val Phe Phe Thr Arg His Asn His50 55 60Lys Ser Pro Thr Asp His Gly Met Leu Gly Glu Trp Cys Asn Gly Asp65 70 75 80
Val Ile Leu Asp Gly Thr Thr Asp Ser Glu Ile Ile Gln Glu Ile Gln85 90 95Gly Gln Val Thr Gly Pro Asp Glu Met Val Glu Lys Asn Thr Tyr Ser100 105 110Ala Phe Asn Lys Thr Arg Leu Gln Glu Asn Leu Glu Lys Ile Gly Val115 120 125Lys Glu Val Ile Val Ile Gly Val Met Thr Asn Leu Cys Cys Glu Thr130 135 140Thr Ala Arg Glu Ala Phe Ile Lys Gly Phe Arg Val Phe Phe Ser Thr145 150 155 160Asp Ala Thr Ala Thr Phe Asn Glu Glu Leu His Glu Ala Thr Leu Met165 170 175Asn Leu Ala Phe Gly Phe Ala Tyr Leu Val Asp Cys Asp Lys Leu Arg180 185 190Arg Ser Leu Leu Gly Asn195<210>13<211>597<212>DNA<213>擬南芥<400>13atggcttctt catcatcgag aacgtacgag acacgaaagc gagagccaaa tcctaaaatc 60gcagctcttc tcgtcatcga tatgcagaat cacttctact ctatggctga accaatcctc120caaaacgctc tcaccaccat cgacatctgc cgacgcgctt caatccccgt attcttcacg180cgccacaacc acaaatcccc aaccgaccac ggcatgctcg gagagtggtg gaacggcgat240ctaatcctcg acggaaccac tgattccgaa atcatcccgg aaatcaatcg ccaggtcacc300ggaccagacg aaatcgtgga gaagagcacg tacagtgcgt ttaacaacac gcaccttcag360gagaagctgg acaagatcgg agtgaaggag gtgatcgtta tcggagtgat gacgaaccta420tgctgtgaga cgacggcgcg tgaagcgttt gtaaaggggt ttagggtttt tttctcgacg480gacgcgactg cgacggttaa tgaagagctt cacgaggcta ctctaatgaa tctcgcgtat540ggctttgctt atctcgtcga ttgcgataga ctccggcgag gtctactcag tagttaa 597<210>14<211>198<212>PRT<213>擬南芥<400>14Met Ala Ser Ser Ser Ser Arg Thr Tyr Glu Thr Arg Lys Arg Glu Pro1 5 10 15Asn Pro Lys Ile Ala Ala Leu Leu Val Ile Asp Met Gln Asn His Phe20 25 30Tyr Ser Met Ala Glu Pro Ile Leu Gln Asn Ala Leu Thr Thr Ile Asp35 40 45Ile Cys Arg Arg Ala Ser Ile Pro Val Phe Phe Thr Arg His Asn His50 55 60Lys Ser Pro Thr Asp His Gly Met Leu Gly Glu Trp Trp Asn Gly Asp65 70 75 80
Leu Ile Leu Asp Gly Thr Thr Asp Ser Glu Ile Ile Pro Glu Ile Asn85 90 95Arg Gln Val Thr Gly Pro Asp Glu Ile Val Glu Lys Ser Thr Tyr Ser100 105 110Ala Phe Asn Asn Thr His Leu Gln Glu Lys Leu Asp Lys Ile Gly Val115 120 125Lys Glu Val Ile Val Ile Gly Val Met Thr Asn Leu Cys Cys Glu Thr130 135 140Thr Ala Arg Glu Ala Phe Val Lys Gly Phe Arg Val Phe Phe Ser Thr145 150 155 160Asp Ala Thr Ala Thr Val Asn Glu Glu Leu His Glu Ala Thr Leu Met165 170 175Asn Leu Ala Tyr Gly Phe Ala Tyr Leu Val Asp Cys Asp Arg Leu Arg180 185 190Arg Gly Leu Leu Ser Ser195<210>15<211>591<212>DNA<213>擬南芥<400>15atggccgaga gatggaggaa cacggctcta ctcgtcatcg acatgcagaa cgatttcata 60gaggaaggtg ctgtgacgca agtgaaagga ggaaaatcta tagttcctaa tgttatcaga120gtcgtcgaac tcgcgaggca gcgtggtatt ctcgtaattt gggttgttcg agaacatgat180cgtcaaggaa gagatgttga attattcagg cgccataact acagttctga gaaagtcggg240ccagttatta aaggcaccgt aggagcagaa ttggttgatg gattgatgat caacgaagaa300gatgactata agattgtgaa aactcgtttc agtgctttct ttagtaccaa tcttcattcc360ttcttgcaaa cttcaggggt taccaagtta gtgattgctg gtgtgcaaac gccgaactgt420atccggcaaa cggtgtttga tgcagtggcg ctggattatc ccaatgtgac tgttattaca480gatgccacag ctgctgcaac accagagatc catactgcga atattcttga catgaagaat540attggagtca agactcctac attacacgag tggtccgaag aacttgcttg a 591<210>16<211>196<212>PRT<213>擬南芥<400>16Met Ala Glu Arg Trp Arg Asn Thr Ala Leu Leu Val Ile Asp Met Gln1 5 10 15Asn Asp Phe Ile Glu Glu Gly Ala Val Thr Gln Val Lys Gly Gly Lys20 25 30Ser Ile Val Pro Asn Val Ile Arg Val Val Glu Leu Ala Arg Gln Arg35 40 45Gly Ile Leu Val Ile Trp Val Val Arg Glu His Asp Arg Gln Gly Arg50 55 60Asp Val Glu Leu Phe Arg Arg His Asn Tyr Ser Ser Glu Lys Val Gly65 70 75 80
Pro Val Ile Lys Gly Thr Val Gly Ala Glu Leu Val Asp Gly Leu Met85 90 95Ile Asn Glu Glu Asp Asp Tyr Lys Ile Val Lys Thr Arg Phe Ser Ala100 105 110Phe Phe Ser Thr Asn Leu His Ser Phe Leu Gln Thr Ser Gly Val Thr115 120 125Lys Leu Val Ile Ala Gly Val Gln Thr Pro Asn Cys Ile Arg Gln Thr130 135 140Val Phe Asp Ala Val Ala Leu Asp Tyr Pro Asn Val Thr Val Ile Thr145 150 155 160Asp Ala Thr Ala Ala Ala Thr Pro Glu Ile His Thr Ala Asn Ile Leu165 170 175Asp Met Lys Asn Ile Gly Val Lys Thr Pro Thr Leu His Glu Trp Ser180 185 190Glu Glu Leu Ala195<210>17<211>1680<212>DNA<213>擬南芥<400>17atggagaaga aagaaaatgg tctcgatgga aagcaatcgg gtcgggtcat taacggaccc 60actaacccga tggtcacacc tctgctcaac gatctttacc aattcaccat ggcttatgct120tattggaaag ctggcaaaca atctgagcga tctgtgtttg atctgtattt tcgtaagaat180ccttttggtg gagaatacac tatctttgct ggtttagaag aatgcatcaa atttctcgct240aatttcaatt tgactgatga agagatcgat ttcgttcgtg attcgttacc tggatgtgag300gaagctttct gtgattatct tcgagggctt gattgttctg acattgaagt gtatgccatt360tcggaaggat cagttgtttt tcctaaagtt cctttactca gaatcgaagg tcctgttgct420gtggtgcaat tgttggaaac tccattcctc aatctcatca attacgcatc tttggttgct480acaaatgcag caagacatcg gtttgttgca ggaaaatcta agcttctgct tgagtttggt540gctagaagag ctcagggacc cgatggtgca ataagcgcat caaagtattg ctaccttgga600ggttttgatg caacaagtaa tgttgcagcg ggaaaactgt ttgggatacc cctccgtggt660actcattccc atgcttttgt tagctcattc atgagccttg atgaaattgt tgacaaagtg720cttcgaagtt ctgatgggaa aagcacttgt aaggatttta tatgtttggt ccaaacttgc780ctaacaaaga ttcagaattc atcttcatta caaggaattt tttccgagac aaatcaaagc840gagcttgcag cgttcatttc atatgcactg gcattcccaa actcctttct cgctcttgta900gacacttatg atgtgatgaa gagtggtatt ccaaacttct gtgctgttgc tctagcactt960aatgaattgg gatacaaagc agtaggcatt agactggatt caggtgactt agcctatctt 1020tctactgagg tcaggaaatt cttttgtgcc atagagagag acctcaaagt tcctgatttc 1080gggaagatga tcgtcactgc tagtaacgat ctaaacgaag agacagtcga tgctctaaat 1140aaacagggtc atgaagtaga tgcatttgga attggaacca acttagtgac ttgctatgcg 1200caagctgcgt taggttgtgt tttcaaactt gtggaaataa acaatcagcc tcggatcaaa 1260ctttctgaag atgttactaa ggtatcgatt ccatgtaaaa agcgtactta cagattgttc 1320ggaaaagagg gttaccctct tgttgatata atgactggag agaacgaacc acctccaaag 1380gtcggtgaaa ggttactttg ccgtcatcca ttcaatgaat caaaaagggc ttatgtggtt 1440ccacaacgcg ttgaagagct tctgaaatgt tattggcgtg gcaatgcaga tgaagctagg 1500gaagagctag agccattgaa agagctaaga aatcgttgca tcaaacagct cgaaaatatg 1560cgacccgatc atatgagaag attaaaccct actccttata aggttagtgt cagcgccaag 1620ttgtatgact tcatccactt cctctggctc aacgaagctc ctgtcggtga actgcattga 1680
<210>18<211>559<212>PRT<213>擬南芥<400>18Met Glu Lys Lys Glu Asn Gly Leu Asp Gly Lys Gln Ser Gly Arg Val1 5 10 15Ile Asn Gly Pro Thr Asn Pro Met Val Thr Pro Leu Leu Asn Asp Leu20 25 30Tyr Gln Phe Thr Met Ala Tyr Ala Tyr Trp Lys Ala Gly Lys Gln Ser35 40 45Glu Arg Ser Val Phe Asp Leu Tyr Phe Arg Lys Asn Pro Phe Gly Gly50 55 60Glu Tyr Thr Ile Phe Ala Gly Leu Glu Glu Cys Ile Lys Phe Leu Ala65 70 75 80Asn Phe Asn Leu Thr Asp Glu Glu Ile Asp Phe Val Arg Asp Ser Leu85 90 95Pro Gly Cys Glu Glu Ala Phe Cys Asp Tyr Leu Arg Gly Leu Asp Cys100 105 110Ser Asp Ile Glu Val Tyr Ala Ile Ser Glu Gly Ser Val Val Phe Pro115 120 125Lys Val Pro Leu Leu Arg Ile Glu Gly Pro Val Ala Val Val Gln Leu130 135 140Leu Glu Thr Pro Phe Leu Asn Leu Ile Asn Tyr Ala Ser Leu Val Ala145 150 155 160Thr Asn Ala Ala Arg His Arg Phe Val Ala Gly Lys Ser Lys Leu Leu165 170 175Leu Glu Phe Gly Ala Arg Arg Ala Gln Gly Pro Asp Gly Ala Ile Ser180 185 190Ala Ser Lys Tyr Cys Tyr Leu Gly Gly Phe Asp Ala Thr Ser Asn Val195 200 205Ala Ala Gly Lys Leu Phe Gly Ile Pro Leu Arg Gly Thr His Ser His210 215 220Ala Phe Val Ser Ser Phe Met Ser Leu Asp Glu Ile Val Asp Lys Val225 230 235 240Leu Arg Ser Ser Asp Gly Lys Ser Thr Cys Lys Asp Phe Ile Cys Leu245 250 255Val Gln Thr Cys Leu Thr Lys Ile Gln Asn Ser Ser Ser Leu Gln Gly260 265 270Ile Phe Ser Glu Thr Asn Gln Ser Glu Leu Ala Ala Phe Ile Ser Tyr275 280 285Ala Leu Ala Phe Pro Asn Ser Phe Leu Ala Leu Val Asp Thr Tyr Asp290 295 300Val Met Lys Ser Gly Ile Pro Asn Phe Cys Ala Val Ala Leu Ala Leu305 310 315 320Asn Glu Leu Gly Tyr Lys Ala Val Gly Ile Arg Leu Asp Ser Gly Asp325 330 335Leu Ala Tyr Leu Ser Thr Glu Val Arg Lys Phe Phe Cys Ala Ile Glu340 345 350
Arg Asp Leu Lys Val Pro Asp Phe Gly Lys Met Ile Val Thr Ala Ser355 360 365Asn Asp Leu Asn Glu Glu Thr Val Asp Ala Leu Asn Lys Gln Gly His370 375 380Glu Val Asp Ala Phe Gly Ile Gly Thr Asn Leu Val Thr Cys Tyr Ala385 390 395 400Gln Ala Ala Leu Gly Cys Val Phe Lys Leu Val Glu Ile Asn Asn Gln405 410 415Pro Arg Ile Lys Leu Ser Glu Asp Val Thr Lys Val Ser Ile Pro Cys420 425 430Lys Lys Arg Thr Tyr Arg Leu Phe Gly Lys Glu Gly Tyr Pro Leu Val435 440 445Asp Ile Met Thr Gly Glu Asn Glu Pro Pro Pro Lys Val Gly Glu Arg450 455 460Leu Leu Cys Arg His Pro Phe Asn Glu Ser Lys Arg Ala Tyr Val Val465 470 475 480Pro Gln Arg Val Glu Glu Leu Leu Lys Cys Tyr Trp Arg Gly Asn Ala485 490 495Asp Glu Ala Arg Glu Glu Leu Glu Pro Leu Lys Glu Leu Arg Asn Arg500 505 510Cys Ile Lys Gln Leu Glu Asn Met Arg Pro Asp His Met Arg Arg Leu515 520 525Asn Pro Thr Pro Tyr Lys Val Ser Val Ser Ala Lys Leu Tyr Asp Phe530 535 540Ile His Phe Leu Trp Leu Asn Glu Ala Pro Val Gly Glu Leu His545 550 555<210>19<211>1674<212>DNA<213>擬南芥<400>19atggagccga aagagaacgg ctcagaattg ggtcagaaga tcattgacgg accaacgaat 60ccaatggtca cacctttact caatgatctt tatcaattca ccatggctta tgcttattgg120aaagctggca aacacaacga acgatccgtt ttcgatctgt attttcgtaa gaacccattt180ggtggtgagt acactgtgtt tgctggatta gaagagtgtg ttaagttctt agccaatttc240aaattgactg atgaagaaat cgatttcgtt caagagtgtt tgcctggatc tgaggaagct300ttttgtgatt atcttagagg gcttgattgt tctgatgttg aagtttatgc aattccggaa360ggatcagttg tttttcctaa agtacctctc atgagagttg aaggacctgt tggtgttgtt420caattgttgg aaactccatt cctcaatctt gtcaattttg catctttggt agctactaac480gcagctaggc atcgctttgt tgccggaaaa tctaagagtc tactcgagtt tggtgctcga540agggctcagg gtccggatgg tgcaataagc gcatcaaaat attgctacct tggaggtttt600gatgcaacaa gtaatgtagc agctggaaaa ctttttggga ttcctcttcg tggaacacac660tctcatgctt atgttagctc attcatgagt actgatgaga ttgttgacaa agtacttcgt720agtgctgatg ggaaaaccac gtgcgaggat tttgttagtc atgttcagac atggttaaaa780aagattcagt attcaccatc tctaagtggc attttctctg agacaaatca aagcgagcta840gcagctttca cctcatatgc actggcattc cccaaaactt ttcttgccct cgtagataca900tacgatgtga tgaagagtgg aatccctaac ttctgtgcag ttgctttagc actcaatgac960tttggatata aagcattagg tattagactg gattcaggtg atttagctta tctatctaga 1020gaggccagaa atttcttctg cacggtagag agagaactaa aagtgcctgg ttttgggaag 1080
atggtcgtca ctgctagtaa tgatctaaat gaagagacga ttgacgcttt aaataaacag1140ggacatgagg tggatgcttt tggcatcggg acctacttgg tcacttgcta ttcacaagcg1200gccttaggtt gcgttttcaa acttgtggag ataaacaatc agcctcggat taaactttct1260gaagatgtta caaaggtatc aataccgtgt aaaaagcgaa gttacagatt atacggcaaa1320gaaggttacc ctctggtaga tataatgact ggagagaacg aaccacctcc aaaggttggt1380gagcgtttac tttgtcgtca cccattcaac gaatccaaaa gagcatatgt agtgccacaa1440cgtgtcgaag agctcctcaa atgttattgg cgtggaagtg cagatgaagc aagagaagta1500ttaccgcctt tgaaagagat aagagaccgt tgcatcaaac agctcgaaaa catgcgacct1560gatcatatga ggagattaaa cccaactcct tataaggtta gtgtaagcgc aaagctgtac1620gatttcatcc acttcttatg gctaaacgaa gcacctgttg gtgaattgca gtga 1674<210>20<211>557<212>PRT<213>擬南芥<400>20Met Glu Pro Lys Glu Asn Gly Ser Glu Leu Gly Gln Lys Ile Ile Asp1 5 10 15Gly Pro Thr Asn Pro Met Val Thr Pro Leu Leu Asn Asp Leu Tyr Gln20 25 30Phe Thr Met Ala Tyr Ala Tyr Trp Lys Ala Gly Lys His Asn Glu Arg35 40 45Ser Val Phe Asp Leu Tyr Phe Arg Lys Asn Pro Phe Gly Gly Glu Tyr50 55 60Thr Val Phe Ala Gly Leu Glu Glu Cys Val Lys Phe Leu Ala Asn Phe65 70 75 80Lys Leu Thr Asp Glu Glu Ile Asp Phe Val Gln Glu Cys Leu Pro Gly85 90 95Ser Glu Glu Ala Phe Cys Asp Tyr Leu Arg Gly Leu Asp Cys Ser Asp100 105 110Val Glu Val Tyr Ala Ile Pro Glu Gly Ser Val Val Phe Pro Lys Val115 120 125Pro Leu Met Arg Val Glu Gly Pro Val Gly Val Val Gln Leu Leu Glu130 135 140Thr Pro Phe Leu Asn Leu Val Asn Phe Ala Ser Leu Val Ala Thr Asn145 150 155 160Ala Ala Arg His Arg Phe Val Ala Gly Lys Ser Lys Ser Leu Leu Glu165 170 175Phe Gly Ala Arg Arg Ala Gln Gly Pro Asp Gly Ala Ile Ser Ala Ser180 185 190Lys Tyr Cys Tyr Leu Gly Gly Phe Asp Ala Thr Ser Asn Val Ala Ala195 200 205Gly Lys Leu Phe Gly Ile Pro Leu Arg Gly Thr His Ser His Ala Tyr210 215 220Val Ser Ser Phe Met Ser Thr Asp Glu Ile Val Asp Lys Val Leu Arg225 230 235 240Ser Ala Asp Gly Lys Thr Thr Cys Glu Asp Phe Val Ser His Val Gln245 250 255Thr Trp Leu Lys Lys Ile Gln Tyr Ser Pro Ser Leu Ser Gly Ile Phe260 265 270
Ser Glu Thr Asn Gln Ser Glu Leu Ala Ala Phe Thr Ser Tyr Ala Leu275 280 285Ala Phe Pro Lys Thr Phe Leu Ala Leu Val Asp Thr Tyr Asp Val Met290 295 300Lys Ser Gly Ile Pro Asn Phe Cys Ala Val Ala Leu Ala Leu Asn Asp305 310 315 320Phe Gly Tyr Lys Ala Leu Gly Ile Arg Leu Asp Ser Gly Asp Leu Ala325 330 335Tyr Leu Ser Arg Glu Ala Arg Asn Phe Phe Cys Thr Val Glu Arg Glu340 345 350Leu Lys Val Pro Gly Phe Gly Lys Met Val Val Thr Ala Ser Asn Asp355 360 365Leu Asn Glu Glu Thr Ile Asp Ala Leu Asn Lys Gln Gly His Glu Val370 375 380Asp Ala Phe Gly Ile Gly Thr Tyr Leu Val Thr Cys Tyr Ser Gln Ala385 390 395 400Ala Leu Gly Cys Val Phe Lys Leu Val Glu Ile Asn Asn Gln Pro Arg405 410 415Ile Lys Leu Ser Glu Asp Val Thr Lys Val Ser Ile Pro Cys Lys Lys420 425 430Arg Ser Tyr Arg Leu Tyr Gly Lys Glu Gly Tyr Pro Leu Val Asp Ile435 440 445Met Thr Gly Glu Asn Glu Pro Pro Pro Lys Val Gly Glu Arg Leu Leu450 455 460Cys Arg His Pro Phe Asn Glu Ser Lys Arg Ala Tyr Val Val Pro Gln465 470 475 480Arg Val Glu Glu Leu Leu Lys Cys Tyr Trp Arg Gly Ser Ala Asp Glu485 490 495Ala Arg Glu Val Leu Pro Pro Leu Lys Glu Ile Arg Asp Arg Cys Ile500 505 510Lys Gln Leu Glu Asn Met Arg Pro Asp His Met Arg Arg Leu Asn Pro515 520 525Thr Pro Tyr Lys Val Ser Val Ser Ala Lys Leu Tyr Asp Phe Ile His530 535 540Phe Leu Trp Leu Asn Glu Ala Pro Val Gly Glu Leu Gln545 550 555<210>21<211>717<212>DNA<213>擬南芥<400>21atggatgtcc cgttaccagt cgagaaatta tcttatggat caaacactga ggacaaaact 60tgtgtagtgc ttgtggcaac tgggagtttc aatcctccta ctttcatgca tttacgcatg120tttgagctgg cgagagatga attacgctca aaaggatttc atgttcttgg aggatatatg180tctcctgtta atgatgcata taagaagaag ggccttttat ctgcagaaca tcgtttagag240atgtgtaatg tatcatgtca aagctctgac tttgtaatgg ttgatccgtg ggaggcatct300caaagcaact accaacgaac tttgacggtt ttatcaaggg tcaagacttt cttaacaaca360aatcgacatg tacccgagga atctctcaaa gtcatgctac tatgtggctc ggatttactg420ctatctttct gcactcccgg tgtttggatc cctgaacagt taagaactat ttgcaaagat480
tatggcattg tgtgcatccg tagagaagga caagatgttg aaaatatgat ctctggtgac540gaaatcttaa acgaaaactg tgctaacgtc aaaatcgttg acaatactgt tcctaatcaa600atcagttcga gtagattaag gcaatgcatt tcgcgagggt tatcggttaa atacttgact660gaagatggag taatagatta tatcagacaa catcaactat acactgagct cacatga 717<210>22<211>238<212>PRT<213>擬南芥<400>22Met Asp Val Pro Leu Pro Val Glu Lys Leu Ser Tyr Gly Ser Asn Thr1 5 10 15Glu Asp Lys Thr Cys Val Val Leu Val Ala Thr Gly Ser Phe Asn Pro20 25 30Pro Thr Phe Met His Leu Arg Met Phe Glu Leu Ala Arg Asp Glu Leu35 40 45Arg Ser Lys Gly Phe His Val Leu Gly Gly Tyr Met Ser Pro Val Asn50 55 60Asp Ala Tyr Lys Lys Lys Gly Leu Leu Ser Ala Glu His Arg Leu Glu65 70 75 80Met Cys Asn Val Ser Cys Gln Ser Ser Asp Phe Val Met Val Asp Pro85 90 95Trp Glu Ala Ser Gln Ser Asn Tyr Gln Arg Thr Leu Thr Val Leu Ser100 105 110Arg Val Lys Thr Phe Leu Thr Thr Asn Arg His Val Pro Glu Glu Ser115 120 125Leu Lys Val Met Leu Leu Cys Gly Ser Asp Leu Leu Leu Ser Phe Cys130 135 140Thr Pro Gly Val Trp Ile Pro Glu Gln Leu Arg Thr Ile Cys Lys Asp145 150 155 160Tyr Gly Ile Val Cys Ile Arg Arg Glu Gly Gln Asp Val Glu Asn Met165 170 175Ile Ser Gly Asp Glu Ile Leu Asn Glu Asn Cys Ala Asn Val Lys Ile180 185 190Val Asp Asn Thr Val Pro Asn Gln Ile Ser Ser Ser Arg Leu Arg Gln195 200 205Cys Ile Ser Arg Gly Leu Ser Val Lys Tyr Leu Thr Glu Asp Gly Val210 215 220Ile Asp Tyr Ile Arg Gln His Gln Leu Tyr Thr Glu Leu Thr225 230 235<210>23<211>2178<212>DNA<213>擬南芥<400>23atgaggctgt tgaaggttgc tacgtgtaac ttgaaccaat gggccatgga tttcgagagc 60aacatgaaga acatcaaggc ttcgatcgct gaggcaaagg ctgctggtgc tgttatcagg120
cttggacccg agctcgaggt cactggctat ggttgcgagg atcacttctt ggaactcgac180actgtcactc atgcgtggga gtgtttgaag gaattgctgc ttggtgattg gacggatgat240attttgtgca gcataggaat gcctgtgatt aaaggagcag agcgttataa ctgccaggtt300ctctgtatga acagaagaat catcatgatt cgaccgaaaa tgtggctcgc aaacgatgga360aactataggg agctacggtg gttcacagct tggaagcaga gagaagagct agaggaattt420cagctcccca ttgaaatttc agaggctttg gagcagaaat cagtcccttt tggttatggt480tacatccagt ttatcgacac ggctgttgca gctgaagtct gtgaggaact gtttagtcca540cttcctcctc atgccgagct cgcattgaat ggtgttgaag tatttatgaa tgcaagtggg600agtcatcacc aacttaggaa actagatatt cgtctgaatg cttttatggg ggctactcat660gctcgtggtg gggtgtatat gtacagtaat caacaaggat gcgatggtag ccgcttatac720tacgatggat gtgcatgtat tgttgtaaac gggaatgttg ttgctcaagg ctcacaattc780tcgttgagag acgttgaggt catcatttca caagtggatc ttgatgcggt tgctagcctt840cgtggatcta taagtagctt tcaggaacaa gcaagctgca aggttaaagt atcttcagta900gctgtgccct gtagacttac acagtccttc aacctgaaaa tgacactaag cagtccgaag960aagatcattt accactctcc acaagaagaa atagcctttg gtcccgcttg ctggatgtgg 1020gactatttga gaagaagtgg cgcttcagga tttttgcttc ctctttctgg cggagcagac 1080agctcctccg tggcagctat tgttggctgc atgtgccaac ttgttgttaa agagattgca 1140aagggagatg agcaagtaaa agctgatgcg aaccgaattg ggaattatgc taatgggcag 1200tttcctactg atagcaaaga gtttgccaaa cgaatatttt acactgtctt tatgggttct 1260gaaaacagtt ctgaggagac aaaaaggcgt tcaaagcagc tggcagacga gattggtgct 1320tggcatcttg atgtttgcat agatggtgtt gtctctgcag ttttatcatt atttcaaaca 1380gttacaggca agcgaccaag gtataaggtt gatggaggat caaatgctga gaaccttggg 1440ttgcagaaca ttcaagcccg gatgagaatg gtgttagcat ttatgttagc gtctctcttg 1500ccttgggttc atagcaaacc aggcttttac cttgttctag gcagctccaa cgttgatgaa 1560ggacttcgtg gttacctgac aaagtatgat tgcagctcag cagacataaa tcctatagga 1620agtatcagta aaatggattt gaggttgttc ttaaaatggg ctgcaacgaa tctcggatat 1680ccatccttgg cagagataga agctgctcca ccaacagctg agcttgagcc cattcgttct 1740gactattctc agctcgatga agtcgacatg ggaatgacat atgaagagct ttcagtctat 1800ggaaggatga ggaagatatt ccgttgtgga ccagtatcta tgttcaagaa tctatgttac 1860aagtggggaa caaagctaag cccagcagaa gtagctgaga aagtgaagta tttcttcaaa 1920tattattcga tcaatcgaca caaaatgact gtcctcacac cgtcttatca cgctgagagt 1980tactccccag aggacaacag attcgatctg aggcagtttc tgtacaacag caagtggcca 2040taccagttta agaagattga cgagattgtt gacagcttaa atggtgactc agttgctttc 2100ccggaagaag aagcaaactc caacaaagaa attggagttg tagcagcaaa ctccggagac 2160ccaagtgcgg gtctctga 2178<210>24<211>725<212>PRT<213>擬南芥<400>24Met Arg Leu Leu Lys Val Ala Thr Cys Asn Leu Asn Gln Trp Ala Met1 5 10 15Asp Phe Glu Ser Asn Met Lys Asn Ile Lys Ala Ser Ile Ala Glu Ala20 25 30Lys Ala Ala Gly Ala Val Ile Arg Leu Gly Pro Glu Leu Glu Val Thr35 40 45Gly Tyr Gly Cys Glu Asp His Phe Leu Glu Leu Asp Thr Val Thr His50 55 60Ala Trp Glu Cys Leu Lys Glu Leu Leu Leu Gly Asp Trp Thr Asp Asp
65 70 75 80Ile Leu Cys Ser Ile Gly Met Pro Val Ile Lys Gly Ala Glu Arg Tyr85 90 95Asn Cys Gln Val Leu Cys Met Asn Arg Arg Ile Ile Met Ile Arg Pro100 105 110Lys Met Trp Leu Ala Asn Asp Gly Asn Tyr Arg Glu Leu Arg Trp Phe115 120 125Thr Ala Trp Lys Gln Arg Glu Glu Leu Glu Glu Phe Gln Leu Pro Ile130 135 140Glu Ile Ser Glu Ala Leu Glu Gln Lys Ser Val Pro Phe Gly Tyr Gly145 150 155 160Tyr Ile Gln Phe Ile Asp Thr Ala Val Ala Ala Glu Val Cys Glu Glu165 170 175Leu Phe Ser Pro Leu Pro Pro His Ala Glu Leu Ala Leu Asn Gly Val180 185 190Glu Val Phe Met Asn Ala Ser Gly Ser His His Gln Leu Arg Lys Leu195 200 205Asp Ile Arg Leu Asn Ala Phe Met Gly Ala Thr His Ala Arg Gly Gly210 215 220Val Tyr Met Tyr Ser Asn Gln Gln Gly Cys Asp Gly Ser Arg Leu Tyr225 230 235 240Tyr Asp Gly Cys Ala Cys Ile Val Val Asn Gly Asn Val Val Ala Gln245 250 255Gly Ser Gln Phe Ser Leu Arg Asp Val Glu Val Ile Ile Ser Gln Val260 265 270Asp Leu Asp Ala Val Ala Ser Leu Arg Gly Ser Ile Ser Ser Phe Gln275 280 285Glu Gln Ala Ser Cys Lys Val Lys Val Ser Ser Val Ala Val Pro Cys290 295 300Arg Leu Thr Gln Ser Phe Asn Leu Lys Met Thr Leu Ser Ser Pro Lys305 310 315 320Lys Ile Ile Tyr His Ser Pro Gln Glu Glu Ile Ala Phe Gly Pro Ala325 330 335Cys Trp Met Trp Asp Tyr Leu Arg Arg Ser Gly Ala Ser Gly Phe Leu340 345 350Leu Pro Leu Ser Gly Gly Ala Asp Ser Ser Ser Val Ala Ala Ile Val355 360 365Gly Cys Met Cys Gln Leu Val Val Lys Glu Ile Ala Lys Gly Asp Glu370 375 380Gln Val Lys Ala Asp Ala Asn Arg Ile Gly Asn Tyr Ala Asn Gly Gln385 390 395 400Phe Pro Thr Asp Ser Lys Glu Phe Ala Lys Arg Ile Phe Tyr Thr Val405 410 415Phe Met Gly Ser Glu Asn Ser Ser Glu Glu Thr Lys Arg Arg Ser Lys420 425 430Gln Leu Ala Asp Glu Ile Gly Ala Trp His Leu Asp Val Cys Ile Asp435 440 445Gly Val Val Ser Ala Val Leu Ser Leu Phe Gln Thr Val Thr Gly Lys450 455 460Arg Pro Arg Tyr Lys Val Asp Gly Gly Ser Asn Ala Glu Asn Leu Gly465 470 475 480
Leu Gln Asn Ile Gln Ala Arg Met Arg Met Val Leu Ala Phe Met Leu485 490 495Ala Ser Leu Leu Pro Trp Val His Ser Lys Pro Gly Phe Tyr Leu Val500 505 510Leu Gly Ser Ser Asn Val Asp Glu Gly Leu Arg Gly Tyr Leu Thr Lys515 520 525Tyr Asp Cys Ser Ser Ala Asp Ile Asn Pro Ile Gly Ser Ile Ser Lys530 535 540Met Asp Leu Arg Leu Phe Leu Lys Trp Ala Ala Thr Asn Leu Gly Tyr545 550 555 560Pro Ser Leu Ala Glu Ile Glu Ala Ala Pro Pro Thr Ala Glu Leu Glu565 570 575Pro Ile Arg Ser Asp Tyr Ser Gln Leu Asp Glu Val Asp Met Gly Met580 585 590Thr Tyr Glu Glu Leu Ser Val Tyr Gly Arg Met Arg Lys Ile Phe Arg595 600 605Cys Gly Pro Val Ser Met Phe Lys Asn Leu Cys Tyr Lys Trp Gly Thr610 615 620Lys Leu Ser Pro Ala Glu Val Ala Glu Lys Val Lys Tyr Phe Phe Lys625 630 635 640Tyr Tyr Ser Ile Asn Arg His Lys Met Thr Val Leu Thr Pro Ser Tyr645 650 655His Ala Glu Ser Tyr Ser Pro Glu Asp Asn Arg Phe Asp Leu Arg Gln660 665 670Phe Leu Tyr Asn Ser Lys Trp Pro Tyr Gln Phe Lys Lys Ile Asp Glu675 680 685Ile Val Asp Ser Leu Asn Gly Asp Ser Val Ala Phe Pro Glu Glu Glu690 695 700Ala Asn Ser Asn Lys Glu Ile Gly Val Val Ala Ala Asn Ser Gly Asp705 710 715 720Pro Ser Ala Gly Leu725<210>25<211>11811<212>DNA<213>人工<220>
<223>pTVE467<400>25agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agcttattta540
tccacgacat tgatgttgtg ggccttcaac tcttccttaa ccttattgat gacttcggga 600tcatcgctga tgggtcttgt gtaatccagc aggacagtgg tcttataacc tagttctgca 660gcggaaatgg cggtggcttt gacacaatac tccaaagcta caccgacaat gtaaacctcg 720tctgtatgat gcttttctaa gtacttgttc atgtcggtct tatggaagtt ccagatgtcg 780tggaaggcgg agtagtattc acggtcagtc aagaaaccct tgtcgacaat cttaatatgc 840ttagtgacca cttggtccat tatttggtca accaattgac taccccaggt gtttttcaca 900cagtgtacgg gccacaaaat accctcttgc gtggaatcat cgcctggcct tggagagtgg 960taggtgtatg ttgaataggg ttctttatct ttatggttct ttgcgaacga aatatgtctg1020gaagggtgcc aatctctggt gaccacaatc ctgtgccagt ctctatcagc atcttgcatc1080aaatccgaga taggattgat taattcctca ccttttggaa cagtcaagga acctaaaggt1140gaaataaaat cattttgcat atcaacaaca attaaagtct ccatggtttt ggtttaataa1200gaagagaaaa gagttctttt gttatggctg aagtaataga gaaatgagct cgagtcctct1260ccaaatgaaa tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg1320cgtcatccct tacgtcagtg gagatatcac atcaatccac ttgctttgaa gacgtggttg1380gaacgtcttc tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt1440cggcagaggc atcttgaacg atagcctttc ctttatcgca atgatggcat ttgtaggtgc1500caccttcctt ttctactgtc cttttgatga agtgacagat agctgggcaa tggaatccga1560ggaggtttcc cgatattacc ctttgttgaa aagtctcaat agccctttgg tcttctgaga1620ctgtatcttt gatattcttg gagtagacga gagtgtcgtg ctccaccatg ttgacgaaga1680ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca gtcttcacgg1740cgagttctgt tagatcctcg atctgaattt ttgactccat gtatggtgca tatggcgcgc1800catatgcccg ggccctgtac agcggccgcg ttaacgcgta tactctagag cgatcgcccg1860ggccggccat ttaaatgaat tcgagctcgg tacccaaacg cggccgcaag ctataacttc1920gtatagcata cattatacga agttattcga ctctagagga tcccaattcc catgcatgga1980gtcaaagatt caaatagagg acacttctcg aactcggccg tcgaactcgg ccgtcgagta2040catggtcgat aagaaaaggc aatttgtaga tgttaattcc catcttgaaa gaaatatagt2100ttaaatattt attgataaaa taacaagtca ggtattatag tccaagcaaa aacataaatt2160tattgatgca agtttaaatt cagaaatatt tcaataactg attatatcag ctggtacatt2220gccgtagatg aaagactgag tgcgatatta tgtgtaatac ataaattgat gatatagcta2280gcttagctca tcgggggatc ctagacgcgt gagatcagat ctcggtgacg ggcaggaccg2340gacggggcgg taccggcagg ctgaagtcca gctgccagaa acccacgtca tgccagttcc2400cgtgcttgaa gccggccgcc cgcagcatgc cgcggggggc atatccgagc gcctcgtgca2460tgcgcacgct cgggtcgttg ggcagcccga tgacagcgac cacgctcttg aagccctgtg2520cctccaggga cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc2580ggggggagac gtacacggtc gactcggccg tccagtcgta ggcgttgcgt gccttccagg2640ggcccgcgta ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc cagggatagc2700gctcccgcag acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga2760agttgaccgt gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg2820cctcggtggc acggcggatg tcggccgggc gtcgttctgg gtccattgtt cttctttact2880ctttgtgtga ctgaggtttg gtctagtgct ttggtcatct atatataatg ataacaacaa2940tgagaacaag ctttggagtg atcggagggt ctaggataca tgagattcaa gtggactagg3000atctacaccg ttggattttg agtgtggata tgtgtgaggt taattttact tggtaacggc3060cacaaaggcc taaggagagg tgttgagacc cttatcggct tgaaccgctg gaataatgcc3120acgtggaaga taattccatg aatcttatcg ttatctatga gtgaaattgt gtgatggtgg3180agtggtgctt gctcatttta cttgcctggt ggacttggcc ctttccttat ggggaattta3240tattttactt actatagagc tttcatacct tttttttacc ttggatttag ttaatatata3300atggtatgat tcatgaataa aaatgggaaa tttttgaatt tgtactgcta aatgcataag3360attaggtgaa actgtggaat atatattttt ttcatttaaa agcaaaattt gccttttact3420agaattataa atatagaaaa atatataaca ttcaaataaa aatgaaaata agaactttca3480aaaaacagaa ctatgtttaa tgtgtaaaga ttagtcgcac atcaagtcat ctgttacaat3540atgttacaac aagtcataag cccaacaaag ttagcacgtc taaataaact aaagagtcca3600cgaaaatatt acaaatcata agcccaacaa agttattgat caaaaaaaaa aaacgcccaa3660
caaagctaaa caaagtccaa aaaaaacttc tcaagtctcc atcttccttt atgaacattg3720aaaactatac acaaaacaag tcagataaat ctctttctgg gcctgtcttc ccaacctcct3780acatcacttc cctatcggat tgaatgtttt acttgtacct tttccgttgc aatgatattg3840atagtatgtt tgtgaaaact aatagggtta acaatcgaag tcatggaata tggatttggt3900ccaagatttt ccgagagctt tctagtagaa agcccatcac cagaaattta ctagtaaaat3960aaatcaccaa ttaggtttct tattatgtgc caaattcaat ataattatag aggatatttc4020aaatgaaaac gtatgaatgt tattagtaaa tggtcaggta agacattaaa aaaatcctac4080gtcagatatt caactttaaa aattcgatca gtgtggaatt gtacaaaaat ttgggatcta4140ctatatatat ataatgcttt acaacacttg gatttttttt tggaggctgg aatttttaat4200ctacatattt gttttggcca tgcaccaact cattgtttag tgtaatactt tgattttgtc4260aaatatatgt gttcgtgtat atttgtataa gaatttcttt gaccatatac acacacacat4320atatatatat atatatatat tatatatcat gcacttttaa ttgaaaaaat aatatatata4380tatatagtgc attttttcta acaaccatat atgttgcgat tgatctgcaa aaatactgct4440agagtaatga aaaatataat ctattgctga aattatctca gatgttaaga ttttcttaaa4500gtaaattctt tcaaatttta gctaaaagtc ttgtaataac taaagaataa tacacaatct4560cgaccacgga aaaaaaacac ataataaatt tgaatttcga ccgcggtacc cggaattggg4620ttataattac ctcaggtcga ggaattaatt cggtacgtac ctaataactt cgtatagcat4680acattatacg aagttatatg gatctcgagg cattacggca ttacggcact cgcgagggtc4740ccaattcgag catggagcca tttacaattg aatatatcct gccgccgctg ccgctttgca4800cccggtggag cttgcatgtt ggtttctacg cagaactgag ccggttaggc agataatttc4860cattgagaac tgagccatgt gcaccttccc cccaacacgg tgagcgacgg ggcaacggag4920tgatccacat gggactttta aacatcatcc gtcggatggc gttgcgagag aagcagtcga4980tccgtgagat cagccgacgc accgggcagg cgcgcaacac gatcgcaaag tatttgaacg5040caggtacaat cgagccgacg ttcacggtac cggaacgacc aagcaagcta gcttagtaaa5100gccctcgcta gattttaatg cggatgttgc gattacttcg ccaactattg cgataacaag5160aaaaagccag cctttcatga tatatctccc aatttgtgta gggcttatta tgcacgctta5220aaaataataa aagcagactt gacctgatag tttggctgtg agcaattatg tgcttagtgc5280atctaacgct tgagttaagc cgcgccgcga agcggcgtcg gcttgaacga attgttagac5340attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact5400gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa5460gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct5520tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat5580ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg5640cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca5700aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg5760ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc5820gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag5880cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag5940ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt6000gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt6060cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca6120ccgcttccct catgatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt6180tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc6240gaggcataga ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc6300cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact6360tgcattacag cttacgaacc gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt6420tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca tttctgtcct6480ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg gcggccttgc6540tgttcttcta cggcaagtgc tgtgcacgga tctgccctgg cttcaggaga tcggaagacc6600tcggccgtcc gggcgcttgc cggtggtgct gaccccggat gaagtctcta gagctctaga6660gggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag cttctgtatg6720gaacgggcat gcggatcagt gagggtttgc aactgcgggt caaggatctg gatttcgatc6780
acggcacgat catcgtgcgg gagggcaagg gctccaagga tcgggccttg atgttacccg6840agagcttggc acccagcctg cgcgagcagg gatcgatcca acccctccgc tgctatagtg6900cagtcggctt ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct6960aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta7020gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg7080ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc7140aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca7200ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct ggcgacgttg7260tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg gacattgccg7320agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg gccgacacca7380ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt7440ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt7500ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag ctgatcgacc7560aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt7620accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct7680tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat gaacgccaag7740aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca ttaccgaaga7800gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc acgtctcaac7860cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg cctggccggc7920cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta7980aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata aacaaatacg8040caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc aggcaagacg8100accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt tctgttagtc8160gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta8220accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat cggccggcgc8280gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc cgcgatcaag8340gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg ggccaccgcc8400gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct acaagcggcc8460tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg8520gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc8580actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag8640gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga ggtaaagaga8700aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg8760ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac tttcagttgc8820cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag accattaccg8880agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga ataaatgagt8940agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac9000gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag cggctgggtt9060gtctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgacggtcg9120caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt9180gaaggccgcg caggccgccc agcggcaacg catcgaggca gaagcacgcc ccggtgaatc9240gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg caaccgccgg cagccggtgc9300gccgtcgatt aggaagccgc ccaagggcga cgagcaacca gattttttcg ttccgatgct9360ctatgacgtg ggcacccgcg atagtcgcag catcatggac gtggccgttt tccgtctgtc9420gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag cttccagacg ggcacgtaga9480ggtttccgca gggccggccg gcatggccag tgtgtgggat tacgacctgg tactgatggc9540ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa gggaagggag acaagcccgg9600ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc tgccggcgag ccgatggcgg9660aaagcagaaa gacgacctgg tagaaacctg cattcggtta aacaccacgc acgttgccat9720gcagcgtacg aagaaggcca agaacggccg cctggtgacg gtatccgagg gtgaagcctt9780gattagccgc tacaagatcg taaagagcga aaccgggcgg ccggagtaca tcgagatcga9840gctagctgat tggatgtacc gcgagatcac agaaggcaag aacccggacg tgctgacggt9900
tcaccccgat tactttttga tcgatcccgg catcggccgt tttctctacc gcctggcacg 9960ccgcgccgca ggcaaggcag aagccagatg gttgttcaag acgatctacg aacgcagtgg10020cagcgccgga gagttcaaga agttctgttt caccgtgcgc aagctgatcg ggtcaaatga10080cctgccggag tacgatttga aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg10140ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg agcagatgct10200agggcaaatt gccctagcag gggaaaaagg tcgaaaaggt ctctttcctg tggatagcac10260gtacattggg aacccaaagc cgtacattgg gaaccggaac ccgtacattg ggaacccaaa10320gccgtacatt gggaaccggt cacacatgta agtgactgat ataaaagaga aaaaaggcga10380tttttccgcc taaaactctt taaaacttat taaaactctt aaaacccgcc tggcctgtgc10440ataactgtct ggccagcgca cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct10500gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat10560ggctggccta cggccaggca atctaccagg gcgcggacaa gccgcgccgt cgccactcga10620ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc10680tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca10740gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc10800agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt10860actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg10920catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg10980gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa11040cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc11100gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc11160aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag11220ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct11280cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta11340ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc11400cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc11460agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt11520gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct11580gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc11640tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca11700agaagatccg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg11760gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc c 11811<210>26<211>11829<212>DNA<213>人工<220>
<223>pTVE468<400>26agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agcttattta540
tccacgacat tgatgttgtg ggccttcaac tcttccttaa ccttattgat gacttcggga 600tcatcgctga tgggtcttgt gtaatccagc aggacagtgg tcttataacc tagttctgca 660gcggaaatgg cggtggcttt gacacaatac tccaaagcta caccgacaat gtaaacctcg 720tctgtatgat gcttttctaa gtacttgttc atgtcggtct tatggaagtt ccagatgtcg 780tggaaggcgg agtagtattc acggtcagtc aagaaaccct tgtcgacaat cttaatatgc 840ttagtgacca cttggtccat tatttggtca accaattgac taccccaggt gtttttcaca 900cagtgtacgg gccacaaaat accctcttgc gtggaatcat cgcctggcct tggagagtgg 960taggtgtatg ttgaataggg ttctttatct ttatggttct ttgcgaacga aatatgtctg1020gaagggtgcc aatctctggt gaccacaatc ctgtgccagt ctctatcagc atcttgcatc1080aaatccgaga taggattgat taattcctca ccttttggaa cagtcaagga acctaaaggt1140gaaataaaat cattttgcat atcaacaacc ttgcgcttct tcttgggaat taaagtctcc1200atggttttgg tttaataaga agagaaaaga gttcttttgt tatggctgaa gtaatagaga1260aatgagctcg agtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg1320aaggatagtg ggattgtgcg tcatccctta cgtcagtgga gatatcacat caatccactt1380gctttgaaga cgtggttgga acgtcttctt tttccacgat gctcctcgtg ggtgggggtc1440catctttggg accactgtcg gcagaggcat cttgaacgat agcctttcct ttatcgcaat1500gatggcattt gtaggtgcca ccttcctttt ctactgtcct tttgatgaag tgacagatag1560ctgggcaatg gaatccgagg aggtttcccg atattaccct ttgttgaaaa gtctcaatag1620ccctttggtc ttctgagact gtatctttga tattcttgga gtagacgaga gtgtcgtgct1680ccaccatgtt gacgaagatt ttcttcttgt cattgagtcg taaaagactc tgtatgaact1740gttcgccagt cttcacggcg agttctgtta gatcctcgat ctgaattttt gactccatgt1800atggtgcata tggcgcgcca tatgcccggg ccctgtacag cggccgcgtt aacgcgtata1860ctctagagcg atcgcccggg ccggccattt aaatgaattc gagctcggta cccaaacgcg1920gccgcaagct ataacttcgt atagcataca ttatacgaag ttattcgact ctagaggatc1980ccaattccca tgcatggagt caaagattca aatagaggac acttctcgaa ctcggccgtc2040gaactcggcc gtcgagtaca tggtcgataa gaaaaggcaa tttgtagatg ttaattccca2100tcttgaaaga aatatagttt aaatatttat tgataaaata acaagtcagg tattatagtc2160caagcaaaaa cataaattta ttgatgcaag tttaaattca gaaatatttc aataactgat2220tatatcagct ggtacattgc cgtagatgaa agactgagtg cgatattatg tgtaatacat2280aaattgatga tatagctagc ttagctcatc gggggatcct agacgcgtga gatcagatct2340cggtgacggg caggaccgga cggggcggta ccggcaggct gaagtccagc tgccagaaac2400ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat2460atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca2520cgctcttgaa gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca2580gtcccgtccg ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg2640cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc gacctcgccg tccacctcgg2700cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt2760cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc2820agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggt2880ccattgttct tctttactct ttgtgtgact gaggtttggt ctagtgcttt ggtcatctat2940atataatgat aacaacaatg agaacaagct ttggagtgat cggagggtct aggatacatg3000agattcaagt ggactaggat ctacaccgtt ggattttgag tgtggatatg tgtgaggtta3060attttacttg gtaacggcca caaaggccta aggagaggtg ttgagaccct tatcggcttg3120aaccgctgga ataatgccac gtggaagata attccatgaa tcttatcgtt atctatgagt3180gaaattgtgt gatggtggag tggtgcttgc tcattttact tgcctggtgg acttggccct3240ttccttatgg ggaatttata ttttacttac tatagagctt tcataccttt tttttacctt3300ggatttagtt aatatataat ggtatgattc atgaataaaa atgggaaatt tttgaatttg3360tactgctaaa tgcataagat taggtgaaac tgtggaatat atattttttt catttaaaag3420caaaatttgc cttttactag aattataaat atagaaaaat atataacatt caaataaaaa3480tgaaaataag aactttcaaa aaacagaact atgtttaatg tgtaaagatt agtcgcacat3540caagtcatct gttacaatat gttacaacaa gtcataagcc caacaaagtt agcacgtcta3600aataaactaa agagtccacg aaaatattac aaatcataag cccaacaaag ttattgatca3660
aaaaaaaaaa acgcccaaca aagctaaaca aagtccaaaa aaaacttctc aagtctccat3720cttcctttat gaacattgaa aactatacac aaaacaagtc agataaatct ctttctgggc3780ctgtcttccc aacctcctac atcacttccc tatcggattg aatgttttac ttgtaccttt3840tccgttgcaa tgatattgat agtatgtttg tgaaaactaa tagggttaac aatcgaagtc3900atggaatatg gatttggtcc aagattttcc gagagctttc tagtagaaag cccatcacca3960gaaatttact agtaaaataa atcaccaatt aggtttctta ttatgtgcca aattcaatat4020aattatagag gatatttcaa atgaaaacgt atgaatgtta ttagtaaatg gtcaggtaag4080acattaaaaa aatcctacgt cagatattca actttaaaaa ttcgatcagt gtggaattgt4140acaaaaattt gggatctact atatatatat aatgctttac aacacttgga tttttttttg4200gaggctggaa tttttaatct acatatttgt tttggccatg caccaactca ttgtttagtg4260taatactttg attttgtcaa atatatgtgt tcgtgtatat ttgtataaga atttctttga4320ccatatacac acacacatat atatatatat atatatatta tatatcatgc acttttaatt4380gaaaaaataa tatatatata tatagtgcat tttttctaac aaccatatat gttgcgattg4440atctgcaaaa atactgctag agtaatgaaa aatataatct attgctgaaa ttatctcaga4500tgttaagatt ttcttaaagt aaattctttc aaattttagc taaaagtctt gtaataacta4560aagaataata cacaatctcg accacggaaa aaaaacacat aataaatttg aatttcgacc4620gcggtacccg gaattgggtt ataattacct caggtcgagg aattaattcg gtacgtacct4680aataacttcg tatagcatac attatacgaa gttatatgga tctcgaggca ttacggcatt4740acggcactcg cgagggtccc aattcgagca tggagccatt tacaattgaa tatatcctgc4800cgccgctgcc gctttgcacc cggtggagct tgcatgttgg tttctacgca gaactgagcc4860ggttaggcag ataatttcca ttgagaactg agccatgtgc accttccccc caacacggtg4920agcgacgggg caacggagtg atccacatgg gacttttaaa catcatccgt cggatggcgt4980tgcgagagaa gcagtcgatc cgtgagatca gccgacgcac cgggcaggcg cgcaacacga5040tcgcaaagta tttgaacgca ggtacaatcg agccgacgtt cacggtaccg gaacgaccaa5100gcaagctagc ttagtaaagc cctcgctaga ttttaatgcg gatgttgcga ttacttcgcc5160aactattgcg ataacaagaa aaagccagcc tttcatgata tatctcccaa tttgtgtagg5220gcttattatg cacgcttaaa aataataaaa gcagacttga cctgatagtt tggctgtgag5280caattatgtg cttagtgcat ctaacgcttg agttaagccg cgccgcgaag cggcgtcggc5340ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt5400ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat5460aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc5520agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg5580acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg5640ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct5700ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca5760gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt5820ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa5880caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca5940aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca6000gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta6060cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag6120tcgatacttc ggcgatcacc gcttccctca tgatgtttaa ctttgtttta gggcgactgc6180cctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg6240cttgctgctt ggatgcccga ggcatagact gtaccccaaa aaaacagtca taacaagcca6300tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc6360gtgagcgcat acgctacttg cattacagct tacgaaccga acaggcttat gtccactggg6420ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag6480tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc6540aggcattggc ggccttgctg ttcttctacg gcaagtgctg tgcacggatc tgccctggct6600tcaggagatc ggaagacctc ggccgtccgg gcgcttgccg gtggtgctga ccccggatga6660agtctctaga gctctagagg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt6720tcgcccagct tctgtatgga acgggcatgc ggatcagtga gggtttgcaa ctgcgggtca6780
aggatctgga tttcgatcac ggcacgatca tcgtgcggga gggcaagggc tccaaggatc6840gggccttgat gttacccgag agcttggcac ccagcctgcg cgagcaggga tcgatccaac6900ccctccgctg ctatagtgca gtcggcttct gacgttcagt gcagccgtct tctgaaaacg6960acatgtcgca caagtcctaa gttacgcgac aggctgccgc cctgcccttt tcctggcgtt7020ttcttgtcgc gtgttttagt cgcataaagt agaatacttg cgactagaac cggagacatt7080acgccatgaa caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg7140accaggactt gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc aagctgtttt7200ccgagaagat caccggcacc aggcgcgacc gcccggagct ggccaggatg cttgaccacc7260tacgccctgg cgacgttgtg acagtgacca ggctagaccg cctggcccgc agcacccgcg7320acctactgga cattgccgag cgcatccagg aggccggcgc gggcctgcgt agcctggcag7380agccgtgggc cgacaccacc acgccggccg gccgcatggt gttgaccgtg ttcgccggca7440ttgccgagtt cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc gaggccgcca7500aggcccgagg cgtgaagttt ggcccccgcc ctaccctcac cccggcacag atcgcgcacg7560cccgcgagct gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg7620tgcatcgctc gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg7680ccaggcggcg cggtgccttc cgtgaggacg cattgaccga ggccgacgcc ctggcggccg7740ccgagaatga acgccaagag gaacaagcat gaaaccgcac caggacggcc aggacgaacc7800gtttttcatt accgaagaga tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc7860gcccgcgcac gtctcaaccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa7920gctggcggcc tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag7980gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg8040agtaaataaa caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa8100ggcgggtcag gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg8160gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg8220cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg8280aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg8340gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac8400gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat8460ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt8520gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag8580cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag8640ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga8700gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga8760gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc8820gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc8880aaggcaagac cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga8940gcaaatgaat aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa9000gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca9060ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca ctggaacccc caagcccgag9120gaatcggcgt gacggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga9180tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga9240agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca9300accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga9360ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt9420ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct9480tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta9540cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg9600gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg9660ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa9720caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt9780atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc9840ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa9900
cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt 9960tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac10020gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa10080gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg10140cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta10200atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggtct10260ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc10320gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat10380aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa10440aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc10500gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc10560cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc10620cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg10680gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt10740aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc10800ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc10860ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg10920cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg10980ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc11040cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag11100gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca11160tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca11220ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg11280atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag11340gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt11400tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca11460cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg11520cggtgctaca gagttcttga agtggtggcc tagctacggc tacactagaa ggacagtatt11580tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc11640cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg11700cagaaaaaaa ggatctcaag aagatccgga aaacgcaagc gcaaagagaa agcaggtagc11760ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc11820ggaattgcc11829<210>27<211>12393<212>DNA<213>人工<220>
<223>pTVE469<400>27agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag480
agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agcttatgct 540atagggaggc ataattgatg cttggcttga ataacagcat cacgatcacc agtatttttc 600atgatatcgt cactaatctt cactgcttta gttccttcag cggaaaaaag cttaataaca 660atattcatag gtttgctcac ttcactggga ttggaaactt tttggaagtc gctggtaaga 720tttgtgccga tgccaaatgc agacttaatg ccacatttct cgcagtattt gtacagttcg 780atacatctgt caacatttaa agcatcgcta tgaacaatta cttttgtgga aggatcgaca 840cctattgatt tatagtgctt tacgactttt tcaatgtatt cctcagcaca accgctatct 900tgacgaacac catggaaaac attggctaaa tcgtcggcag aattggctgt aaaagatttg 960agaaacacat cagtagagaa tgtatccgtt aaggctatta aaagactagt accaaaagtt1020tggacccact ttaaggaagc aatacgattt gcttgtttat aattttgagt aatagctgca1080atgcccatat accactcgtg agcaaccgta ccagagacat ttagattata tttggcggcg1140aagtaaacat tagatgtacc aaggaaactt ccagggccct taaaatcctc ttgtgctttc1200atgagacctt ggagaacaat ttcctgggtg tgaggatcac gacgacgacg agtgccaaag1260tcagtaaagg cacatccggc tcggatgaga cgcttaccct tctcgtaagc tttttcaaac1320tgaccctcag gtgaccagtc cttatcgaca aatttaaaat aagattctga gacgagagca1380agcagtggaa tttcataaaa aatggtattc ttccagaggc cgtgaataaa gattgagaga1440tccttagttt cagaatcata attaagggaa attgaatttt caggatcaaa ttcgaactca1500tgcatgaatt cataaaatga ttcctttaaa taaggacagt tcttgcgaag ccattgctct1560tcttcaggaa gtaaatgtaa attccgtaag cctcttattt gttcccgtaa ccagttataa1620gcctcctgat ttaatgccat ttttggggac cggtttgtat acttatatga tacttgagca1680tccggataat gctctaaaac ggcttgaagc atggtgagtt tgtaaagatc cgtatcgagg1740atagagacaa cagccggttc acccatggtt ttggtttaat aagaagagaa aagagttctt1800ttgttatggc tgaagtaata gagaaatgag ctcgagtcct ctccaaatga aatgaacttc1860cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag1920tggagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca1980cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa2040cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg2100tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta2160ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct2220tggagtagac gagagtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga2280gtcgtaaaag actctgtatg aactgttcgc cagtcttcac ggcgagttct gttagatcct2340cgatctgaat ttttgactcc atgtatggtg catatggcgc gccatatgcc cgggccctgt2400acagcggccg cgttaacgcg tatactctag agcgatcgcc cgggccggcc atttaaatga2460attcgagctc ggtacccaaa cgcggccgca agctataact tcgtatagca tacattatac2520gaagttattc gactctagag gatcccaatt cccatgcatg gagtcaaaga ttcaaataga2580ggacacttct cgaactcggc cgtcgaactc ggccgtcgag tacatggtcg ataagaaaag2640gcaatttgta gatgttaatt cccatcttga aagaaatata gtttaaatat ttattgataa2700aataacaagt caggtattat agtccaagca aaaacataaa tttattgatg caagtttaaa2760ttcagaaata tttcaataac tgattatatc agctggtaca ttgccgtaga tgaaagactg2820agtgcgatat tatgtgtaat acataaattg atgatatagc tagcttagct catcggggga2880tcctagacgc gtgagatcag atctcggtga cgggcaggac cggacggggc ggtaccggca2940ggctgaagtc cagctgccag aaacccacgt catgccagtt cccgtgcttg aagccggccg3000cccgcagcat gccgcggggg gcatatccga gcgcctcgtg catgcgcacg ctcgggtcgt3060tgggcagccc gatgacagcg accacgctct tgaagccctg tgcctccagg gacttcagca3120ggtgggtgta gagcgtggag cccagtcccg tccgctggtg gcggggggag acgtacacgg3180tcgactcggc cgtccagtcg taggcgttgc gtgccttcca ggggcccgcg taggcgatgc3240cggcgacctc gccgtccacc tcggcgacga gccagggata gcgctcccgc agacggacga3300ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg gaagttgacc gtgcttgtct3360cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg gcacggcgga3420tgtcggccgg gcgtcgttct gggtccattg ttcttcttta ctctttgtgt gactgaggtt3480tggtctagtg ctttggtcat ctatatataa tgataacaac aatgagaaca agctttggag3540tgatcggagg gtctaggata catgagattc aagtggacta ggatctacac cgttggattt3600
tgagtgtgga tatgtgtgag gttaatttta cttggtaacg gccacaaagg cctaaggaga3660ggtgttgaga cccttatcgg cttgaaccgc tggaataatg ccacgtggaa gataattcca3720tgaatcttat cgttatctat gagtgaaatt gtgtgatggt ggagtggtgc ttgctcattt3780tacttgcctg gtggacttgg ccctttcctt atggggaatt tatattttac ttactataga3840gctttcatac ctttttttta ccttggattt agttaatata taatggtatg attcatgaat3900aaaaatggga aatttttgaa tttgtactgc taaatgcata agattaggtg aaactgtgga3960atatatattt ttttcattta aaagcaaaat ttgcctttta ctagaattat aaatatagaa4020aaatatataa cattcaaata aaaatgaaaa taagaacttt caaaaaacag aactatgttt4080aatgtgtaaa gattagtcgc acatcaagtc atctgttaca atatgttaca acaagtcata4140agcccaacaa agttagcacg tctaaataaa ctaaagagtc cacgaaaata ttacaaatca4200taagcccaac aaagttattg atcaaaaaaa aaaaacgccc aacaaagcta aacaaagtcc4260aaaaaaaact tctcaagtct ccatcttcct ttatgaacat tgaaaactat acacaaaaca4320agtcagataa atctctttct gggcctgtct tcccaacctc ctacatcact tccctatcgg4380attgaatgtt ttacttgtac cttttccgtt gcaatgatat tgatagtatg tttgtgaaaa4440ctaatagggt taacaatcga agtcatggaa tatggatttg gtccaagatt ttccgagagc4500tttctagtag aaagcccatc accagaaatt tactagtaaa ataaatcacc aattaggttt4560cttattatgt gccaaattca atataattat agaggatatt tcaaatgaaa acgtatgaat4620gttattagta aatggtcagg taagacatta aaaaaatcct acgtcagata ttcaacttta4680aaaattcgat cagtgtggaa ttgtacaaaa atttgggatc tactatatat atataatgct4740ttacaacact tggatttttt tttggaggct ggaattttta atctacatat ttgttttggc4800catgcaccaa ctcattgttt agtgtaatac tttgattttg tcaaatatat gtgttcgtgt4860atatttgtat aagaatttct ttgaccatat acacacacac atatatatat atatatatat4920attatatatc atgcactttt aattgaaaaa ataatatata tatatatagt gcattttttc4980taacaaccat atatgttgcg attgatctgc aaaaatactg ctagagtaat gaaaaatata5040atctattgct gaaattatct cagatgttaa gattttctta aagtaaattc tttcaaattt5100tagctaaaag tcttgtaata actaaagaat aatacacaat ctcgaccacg gaaaaaaaac5160acataataaa tttgaatttc gaccgcggta cccggaattg ggttataatt acctcaggtc5220gaggaattaa ttcggtacgt acctaataac ttcgtatagc atacattata cgaagttata5280tggatctcga ggcattacgg cattacggca ctcgcgaggg tcccaattcg agcatggagc5340catttacaat tgaatatatc ctgccgccgc tgccgctttg cacccggtgg agcttgcatg5400ttggtttcta cgcagaactg agccggttag gcagataatt tccattgaga actgagccat5460gtgcaccttc cccccaacac ggtgagcgac ggggcaacgg agtgatccac atgggacttt5520taaacatcat ccgtcggatg gcgttgcgag agaagcagtc gatccgtgag atcagccgac5580gcaccgggca ggcgcgcaac acgatcgcaa agtatttgaa cgcaggtaca atcgagccga5640cgttcacggt accggaacga ccaagcaagc tagcttagta aagccctcgc tagattttaa5700tgcggatgtt gcgattactt cgccaactat tgcgataaca agaaaaagcc agcctttcat5760gatatatctc ccaatttgtg tagggcttat tatgcacgct taaaaataat aaaagcagac5820ttgacctgat agtttggctg tgagcaatta tgtgcttagt gcatctaacg cttgagttaa5880gccgcgccgc gaagcggcgt cggcttgaac gaattgttag acattatttg ccgactacct5940tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca6000agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact6060gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg6120ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc6180agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt6240caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc6300ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg6360caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc6420acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct6480ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat6540caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat6600ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga6660cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatgatgt6720
ttaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca6780aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc6840caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt6900cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa6960ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc7020cggcaacctt gggcagcagc gaagtcgagg catttctgtc ctggctggcg aacgagcgca7080aggtttcggt ctccacgcat cgtcaggcat tggcggcctt gctgttcttc tacggcaagt7140gctgtgcacg gatctgccct ggcttcagga gatcggaaga cctcggccgt ccgggcgctt7200gccggtggtg ctgaccccgg atgaagtctc tagagctcta gagggttcgc atcctcggtt7260ttctggaagg cgagcatcgt ttgttcgccc agcttctgta tggaacgggc atgcggatca7320gtgagggttt gcaactgcgg gtcaaggatc tggatttcga tcacggcacg atcatcgtgc7380gggagggcaa gggctccaag gatcgggcct tgatgttacc cgagagcttg gcacccagcc7440tgcgcgagca gggatcgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt7500cagtgcagcc gtcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg7560ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata7620cttgcgacta gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg7680gctatgcccg cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg7740cggccggctg caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg7800agctggccag gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag7860accgcctggc ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg7920gcgcgggcct gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca7980tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca8040cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc8100tcaccccggc acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga8160aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca8220gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga8280ccgaggccga cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc8340gcaccaggac ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat8400cgcggccggg tacgtgttcg agccgcccgc gcacgtctca accgtgcggc tgcatgaaat8460cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga8520aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc8580ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag8640gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta8700gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc8760agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac8820cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac8880ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg8940attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt9000aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg9060atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc9120attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca9180accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct9240gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa9300acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc9360tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca9420agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca9480tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct9540aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg9600tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg gccctgcaat9660ggcactggaa cccccaagcc cgaggaatcg gcgtgacggt cgcaaaccat ccggcccggt9720acaaatcggc gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc9780ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga9840
tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc 9900gcccaagggc gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg 9960cgatagtcgc agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc10020tggcgaggtg atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc10080cggcatggcc agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga10140atccatgaac cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca10200cgttgcggac gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct10260ggtagaaacc tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc10320caagaacggc cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat10380cgtaaagagc gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta10440ccgcgagatc acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt10500gatcgatccc ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc10560agaagccaga tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa10620gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt10680gaaggaggag gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga10740gggcgaagca tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc10800aggggaaaaa ggtcgaaaag gtctctttcc tgtggatagc acgtacattg ggaacccaaa10860gccgtacatt gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg10920gtcacacatg taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc10980tttaaaactt attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg11040cacagccgaa gagctgcaaa aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc11100cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa atggctggcc tacggccagg11160caatctacca gggcgcggac aagccgcgcc gtcgccactc gaccgccggc gcccacatca11220aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc11280cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg11340cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg11400gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat11460gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc11520ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca11580ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg11640agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca11700taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa11760cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc11820tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc11880gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct11940gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg12000tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag12060gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta12120cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg12180aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt12240tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc cggaaaacgc12300aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc12360ggttttatgg acagcaagcg aaccggaatt gcc 12393<210>28<211>12414<212>DNA<213>人工<220>
<223>pTVE 470
<400>28agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg 120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga 180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg 240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat 300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt 360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat 420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag 480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agcttatgct 540atagggaggc ataattgatg cttggcttga ataacagcat cacgatcacc agtatttttc 600atgatatcgt cactaatctt cactgcttta gttccttcag cggaaaaaag cttaataaca 660atattcatag gtttgctcac ttcactggga ttggaaactt tttggaagtc gctggtaaga 720tttgtgccga tgccaaatgc agacttaatg ccacatttct cgcagtattt gtacagttcg 780atacatctgt caacatttaa agcatcgcta tgaacaatta cttttgtgga aggatcgaca 840cctattgatt tatagtgctt tacgactttt tcaatgtatt cctcagcaca accgctatct 900tgacgaacac catggaaaac attggctaaa tcgtcggcag aattggctgt aaaagatttg 960agaaacacat cagtagagaa tgtatccgtt aaggctatta aaagactagt accaaaagtt1020tggacccact ttaaggaagc aatacgattt gcttgtttat aattttgagt aatagctgca1080atgcccatat accactcgtg agcaaccgta ccagagacat ttagattata tttggcggcg1140aagtaaacat tagatgtacc aaggaaactt ccagggccct taaaatcctc ttgtgctttc1200atgagacctt ggagaacaat ttcctgggtg tgaggatcac gacgacgacg agtgccaaag1260tcagtaaagg cacatccggc tcggatgaga cgcttaccct tctcgtaagc tttttcaaac1320tgaccctcag gtgaccagtc cttatcgaca aatttaaaat aagattctga gacgagagca1380agcagtggaa tttcataaaa aatggtattc ttccagaggc cgtgaataaa gattgagaga1440tccttagttt cagaatcata attaagggaa attgaatttt caggatcaaa ttcgaactca1500tgcatgaatt cataaaatga ttcctttaaa taaggacagt tcttgcgaag ccattgctct1560tcttcaggaa gtaaatgtaa attccgtaag cctcttattt gttcccgtaa ccagttataa1620gcctcctgat ttaatgccat ttttggggac cggtttgtat acttatatga tacttgagca1680tccggataat gctctaaaac ggcttgaagc atggtgagtt tgtaaagatc cgtatcgagg1740atagagacaa cagcaacctt gcgcttcttc ttgggcggtt cacccatggt tttggtttaa1800taagaagaga aaagagttct tttgttatgg ctgaagtaat agagaaatga gctcgagtcc1860tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga tagtgggatt1920gtgcgtcatc ccttacgtca gtggagatat cacatcaatc cacttgcttt gaagacgtgg1980ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac2040tgtcggcaga ggcatcttga acgatagcct ttcctttatc gcaatgatgg catttgtagg2100tgccaccttc cttttctact gtccttttga tgaagtgaca gatagctggg caatggaatc2160cgaggaggtt tcccgatatt accctttgtt gaaaagtctc aatagccctt tggtcttctg2220agactgtatc tttgatattc ttggagtaga cgagagtgtc gtgctccacc atgttgacga2280agattttctt cttgtcattg agtcgtaaaa gactctgtat gaactgttcg ccagtcttca2340cggcgagttc tgttagatcc tcgatctgaa tttttgactc catgtatggt gcatatggcg2400cgccatatgc ccgggccctg tacagcggcc gcgttaacgc gtatactcta gagcgatcgc2460ccgggccggc catttaaatg aattcgagct cggtacccaa acgcggccgc aagctataac2520ttcgtatagc atacattata cgaagttatt cgactctaga ggatcccaat tcccatgcat2580ggagtcaaag attcaaatag aggacacttc tcgaactcgg ccgtcgaact cggccgtcga2640gtacatggtc gataagaaaa ggcaatttgt agatgttaat tcccatcttg aaagaaatat2700agtttaaata tttattgata aaataacaag tcaggtatta tagtccaagc aaaaacataa2760atttattgat gcaagtttaa attcagaaat atttcaataa ctgattatat cagctggtac2820attgccgtag atgaaagact gagtgcgata ttatgtgtaa tacataaatt gatgatatag2880ctagcttagc tcatcggggg atcctagacg cgtgagatca gatctcggtg acgggcagga2940ccggacgggg cggtaccggc aggctgaagt ccagctgcca gaaacccacg tcatgccagt3000
tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg ggcatatccg agcgcctcgt3060gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc gaccacgctc ttgaagccct3120gtgcctccag ggacttcagc aggtgggtgt agagcgtgga gcccagtccc gtccgctggt3180ggcgggggga gacgtacacg gtcgactcgg ccgtccagtc gtaggcgttg cgtgccttcc3240aggggcccgc gtaggcgatg ccggcgacct cgccgtccac ctcggcgacg agccagggat3300agcgctcccg cagacggacg aggtcgtccg tccactcctg cggttcctgc ggctcggtac3360ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat ggtgcagacc gccggcatgt3420ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc tgggtccatt gttcttcttt3480actctttgtg tgactgaggt ttggtctagt gctttggtca tctatatata atgataacaa3540caatgagaac aagctttgga gtgatcggag ggtctaggat acatgagatt caagtggact3600aggatctaca ccgttggatt ttgagtgtgg atatgtgtga ggttaatttt acttggtaac3660ggccacaaag gcctaaggag aggtgttgag acccttatcg gcttgaaccg ctggaataat3720gccacgtgga agataattcc atgaatctta tcgttatcta tgagtgaaat tgtgtgatgg3780tggagtggtg cttgctcatt ttacttgcct ggtggacttg gccctttcct tatggggaat3840ttatatttta cttactatag agctttcata cctttttttt accttggatt tagttaatat3900ataatggtat gattcatgaa taaaaatggg aaatttttga atttgtactg ctaaatgcat3960aagattaggt gaaactgtgg aatatatatt tttttcattt aaaagcaaaa tttgcctttt4020actagaatta taaatataga aaaatatata acattcaaat aaaaatgaaa ataagaactt4080tcaaaaaaca gaactatgtt taatgtgtaa agattagtcg cacatcaagt catctgttac4140aatatgttac aacaagtcat aagcccaaca aagttagcac gtctaaataa actaaagagt4200ccacgaaaat attacaaatc ataagcccaa caaagttatt gatcaaaaaa aaaaaacgcc4260caacaaagct aaacaaagtc caaaaaaaac ttctcaagtc tccatcttcc tttatgaaca4320ttgaaaacta tacacaaaac aagtcagata aatctctttc tgggcctgtc ttcccaacct4380cctacatcac ttccctatcg gattgaatgt tttacttgta ccttttccgt tgcaatgata4440ttgatagtat gtttgtgaaa actaataggg ttaacaatcg aagtcatgga atatggattt4500ggtccaagat tttccgagag ctttctagta gaaagcccat caccagaaat ttactagtaa4560aataaatcac caattaggtt tcttattatg tgccaaattc aatataatta tagaggatat4620ttcaaatgaa aacgtatgaa tgttattagt aaatggtcag gtaagacatt aaaaaaatcc4680tacgtcagat attcaacttt aaaaattcga tcagtgtgga attgtacaaa aatttgggat4740ctactatata tatataatgc tttacaacac ttggattttt ttttggaggc tggaattttt4800aatctacata tttgttttgg ccatgcacca actcattgtt tagtgtaata ctttgatttt4860gtcaaatata tgtgttcgtg tatatttgta taagaatttc tttgaccata tacacacaca4920catatatata tatatatata tattatatat catgcacttt taattgaaaa aataatatat4980atatatatag tgcatttttt ctaacaacca tatatgttgc gattgatctg caaaaatact5040gctagagtaa tgaaaaatat aatctattgc tgaaattatc tcagatgtta agattttctt5100aaagtaaatt ctttcaaatt ttagctaaaa gtcttgtaat aactaaagaa taatacacaa5160tctcgaccac ggaaaaaaaa cacataataa atttgaattt cgaccgcggt acccggaatt5220gggttataat tacctcaggt cgaggaatta attcggtacg tacctaataa cttcgtatag5280catacattat acgaagttat atggatctcg aggcattacg gcattacggc actcgcgagg5340gtcccaattc gagcatggag ccatttacaa ttgaatatat cctgccgccg ctgccgcttt5400gcacccggtg gagcttgcat gttggtttct acgcagaact gagccggtta ggcagataat5460ttccattgag aactgagcca tgtgcacctt ccccccaaca cggtgagcga cggggcaacg5520gagtgatcca catgggactt ttaaacatca tccgtcggat ggcgttgcga gagaagcagt5580cgatccgtga gatcagccga cgcaccgggc aggcgcgcaa cacgatcgca aagtatttga5640acgcaggtac aatcgagccg acgttcacgg taccggaacg accaagcaag ctagcttagt5700aaagccctcg ctagatttta atgcggatgt tgcgattact tcgccaacta ttgcgataac5760aagaaaaagc cagcctttca tgatatatct cccaatttgt gtagggctta ttatgcacgc5820ttaaaaataa taaaagcaga cttgacctga tagtttggct gtgagcaatt atgtgcttag5880tgcatctaac gcttgagtta agccgcgccg cgaagcggcg tcggcttgaa cgaattgtta5940gacattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca6000actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt6060caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat6120
ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta6180catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta6240gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta6300ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg6360tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt6420cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta6480cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca6540aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac6600tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg6660gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga6720tcaccgcttc cctcatgatg tttaactttg ttttagggcg actgccctgc tgcgtaacat6780cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg6840cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa accgccactg6900cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct6960acttgcatta cagcttacga accgaacagg cttatgtcca ctgggttcgt gccttcatcc7020gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt7080cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct7140tgctgttctt ctacggcaag tgctgtgcac ggatctgccc tggcttcagg agatcggaag7200acctcggccg tccgggcgct tgccggtggt gctgaccccg gatgaagtct ctagagctct7260agagggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc cagcttctgt7320atggaacggg catgcggatc agtgagggtt tgcaactgcg ggtcaaggat ctggatttcg7380atcacggcac gatcatcgtg cgggagggca agggctccaa ggatcgggcc ttgatgttac7440ccgagagctt ggcacccagc ctgcgcgagc agggatcgat ccaacccctc cgctgctata7500gtgcagtcgg cttctgacgt tcagtgcagc cgtcttctga aaacgacatg tcgcacaagt7560cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt7620ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga7680gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca7740accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg7800gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg7860ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg7920ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca7980ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc8040gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga8100agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg8160accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc8220tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg8280ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc8340aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga8400agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacgtctc8460aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc8520ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga8580gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat8640acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag8700acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta8760gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg8820ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg8880cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc8940aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc9000gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg9060gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg9120ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca9180ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc9240
gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag 9300agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca 9360aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt 9420tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta 9480ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg 9540agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc 9600gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg 9660gttgtctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgacgg 9720tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa 9780gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga 9840atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg 9900tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat 9960gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct10020gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt10080agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat10140ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc10200cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg10260cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc10320catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc10380cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat10440cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac10500ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc10560acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag10620tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa10680tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat10740gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat10800gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggtctctttc ctgtggatag10860cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc10920aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg10980cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg11040tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc11100gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa11160aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact11220cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa11280acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga11340gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga11400cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat11460tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata11520ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct11580gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga11640taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc11700cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg11760ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg11820aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt11880tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt11940gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg12000cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact12060ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt12120cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct12180gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac12240cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc12300tcaagaagat ccggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac12360
atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgcc 12414<210>29<211>12366<212>DNA<213>人工<220>
<223>pTVE496<400>29agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg 120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga 180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg 240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat 300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt 360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat 420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag 480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agctcattct 540ttgtttccaa gaacttgctt aacaggttcg gtttggtcca catatagtct atgttcttgg 600atatacctga tgaccgaatt aggtaacaaa tattgtacag acatggcgcg tctgataaat 660agacgaactt tcgtggaaga aatatcatta tagatgagtt gcttgatgat aagaatattc 720cttctatgtt catacataat atcatgggat aacaaaaaag accttacatc agaaccagta 780cgttcgacaa tcaaacaacc gtaattaccg agaatgtgat gtaaatcggc gtccgcccaa 840acgtttggtt cacccattga ctctattagg tcaccaccag ccagcaacat tattttcaca 900ccaatttttt ctccagtaac agtagctaca ccacctctct taatattgat ttcgtgattg 960aaatgatcca agaccttggc agttcttgtg tatgaaggtt gcaatgactc ccatgcatcc1020accatcaacc aagatgaggt tctttcgcag gccaattcac acatacgtac tctatggtag1080gatggggcca agccttgctt ttgatagtta tcactaacag gggagtaata tccacctatg1140acttcaaacc ttgtttgttc agagattgca tctaaagcca tttcaaacat tcttagatgc1200aagtaggtga ttggtgaaaa agacccacat gctactatta ctaacggcag tttatttgga1260tccagtaatt ttttcgataa tctgtgtgag gggaattcgt agtcttccaa ggttcttgct1320tgacgaacga ttccatgtgg aacttcttct aaatcagcaa tctgatattt ctgaacacct1380ctagtttctg attctgtggt tgaatctgaa ccgtatctct ccttggatct aaaatccgcg1440tcatcatctt cggaagacac ttctgcggaa agtggttgaa agtcgtcttg gttcaatgga1500atatgctgat gttttttatc attgccttct ttacgactgt gatgatggtg atgatgatgc1560ttaggatgct ttttcttcct cttaatatta aaaggtgcat ctatagagga attcgcatcg1620gctaagacgt atggaataat tggaatagat ttgggaattt tagattccgg gtcgggtgga1680ggaatcaatt cctcgtctgc agatggcggt ttgaaatccg gagctcttgt gggatccatg1740gttttggttt aataagaaga gaaaagagtt cttttgttat ggctgaagta atagagaaat1800gagctcgagt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag1860gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct1920ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat1980ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat2040ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg2100ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc2160tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg tcgtgctcca2220ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt2280cgccagtctt cacggcgagt tctgttagat cctcgatctg aatttttgac tccatgtatg2340gtgcatatgg cgcgccatat gcccgggccc tgtacagcgg ccgcgttaac gcgtatactc2400
tagagcgatc gcccgggccg gccatttaaa tgaattcgag ctcggtaccc aaacgcggcc2460gcaagctata acttcgtata gcatacatta tacgaagtta ttcgactcta gaggatccca2520attcccatgc atggagtcaa agattcaaat agaggacact tctcgaactc ggccgtcgaa2580ctcggccgtc gagtacatgg tcgataagaa aaggcaattt gtagatgtta attcccatct2640tgaaagaaat atagtttaaa tatttattga taaaataaca agtcaggtat tatagtccaa2700gcaaaaacat aaatttattg atgcaagttt aaattcagaa atatttcaat aactgattat2760atcagctggt acattgccgt agatgaaaga ctgagtgcga tattatgtgt aatacataaa2820ttgatgatat agctagctta gctcatcggg ggatcctaga cgcgtgagat cagatctcgg2880tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca2940cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc3000cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc3060tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc3120ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt3180tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga3240cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct3300gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga3360ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggtcca3420ttgttcttct ttactctttg tgtgactgag gtttggtcta gtgctttggt catctatata3480taatgataac aacaatgaga acaagctttg gagtgatcgg agggtctagg atacatgaga3540ttcaagtgga ctaggatcta caccgttgga ttttgagtgt ggatatgtgt gaggttaatt3600ttacttggta acggccacaa aggcctaagg agaggtgttg agacccttat cggcttgaac3660cgctggaata atgccacgtg gaagataatt ccatgaatct tatcgttatc tatgagtgaa3720attgtgtgat ggtggagtgg tgcttgctca ttttacttgc ctggtggact tggccctttc3780cttatgggga atttatattt tacttactat agagctttca tacctttttt ttaccttgga3840tttagttaat atataatggt atgattcatg aataaaaatg ggaaattttt gaatttgtac3900tgctaaatgc ataagattag gtgaaactgt ggaatatata tttttttcat ttaaaagcaa3960aatttgcctt ttactagaat tataaatata gaaaaatata taacattcaa ataaaaatga4020aaataagaac tttcaaaaaa cagaactatg tttaatgtgt aaagattagt cgcacatcaa4080gtcatctgtt acaatatgtt acaacaagtc ataagcccaa caaagttagc acgtctaaat4140aaactaaaga gtccacgaaa atattacaaa tcataagccc aacaaagtta ttgatcaaaa4200aaaaaaaacg cccaacaaag ctaaacaaag tccaaaaaaa acttctcaag tctccatctt4260cctttatgaa cattgaaaac tatacacaaa acaagtcaga taaatctctt tctgggcctg4320tcttcccaac ctcctacatc acttccctat cggattgaat gttttacttg taccttttcc4380gttgcaatga tattgatagt atgtttgtga aaactaatag ggttaacaat cgaagtcatg4440gaatatggat ttggtccaag attttccgag agctttctag tagaaagccc atcaccagaa4500atttactagt aaaataaatc accaattagg tttcttatta tgtgccaaat tcaatataat4560tatagaggat atttcaaatg aaaacgtatg aatgttatta gtaaatggtc aggtaagaca4620ttaaaaaaat cctacgtcag atattcaact ttaaaaattc gatcagtgtg gaattgtaca4680aaaatttggg atctactata tatatataat gctttacaac acttggattt ttttttggag4740gctggaattt ttaatctaca tatttgtttt ggccatgcac caactcattg tttagtgtaa4800tactttgatt ttgtcaaata tatgtgttcg tgtatatttg tataagaatt tctttgacca4860tatacacaca cacatatata tatatatata tatattatat atcatgcact tttaattgaa4920aaaataatat atatatatat agtgcatttt ttctaacaac catatatgtt gcgattgatc4980tgcaaaaata ctgctagagt aatgaaaaat ataatctatt gctgaaatta tctcagatgt5040taagattttc ttaaagtaaa ttctttcaaa ttttagctaa aagtcttgta ataactaaag5100aataatacac aatctcgacc acggaaaaaa aacacataat aaatttgaat ttcgaccgcg5160gtacccggaa ttgggttata attacctcag gtcgaggaat taattcggta cgtacctaat5220aacttcgtat agcatacatt atacgaagtt atatggatct cgaggcatta cggcattacg5280gcactcgcga gggtcccaat tcgagcatgg agccatttac aattgaatat atcctgccgc5340cgctgccgct ttgcacccgg tggagcttgc atgttggttt ctacgcagaa ctgagccggt5400taggcagata atttccattg agaactgagc catgtgcacc ttccccccaa cacggtgagc5460gacggggcaa cggagtgatc cacatgggac ttttaaacat catccgtcgg atggcgttgc5520
gagagaagca gtcgatccgt gagatcagcc gacgcaccgg gcaggcgcgc aacacgatcg5580caaagtattt gaacgcaggt acaatcgagc cgacgttcac ggtaccggaa cgaccaagca5640agctagctta gtaaagccct cgctagattt taatgcggat gttgcgatta cttcgccaac5700tattgcgata acaagaaaaa gccagccttt catgatatat ctcccaattt gtgtagggct5760tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa5820ttatgtgctt agtgcatcta acgcttgagt taagccgcgc cgcgaagcgg cgtcggcttg5880aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga5940caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag6000cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt6060cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca6120acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta6180aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg6240ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat6300caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc6360caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa6420tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa6480ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca6540aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg6600ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg6660atacttcggc gatcaccgct tccctcatga tgtttaactt tgttttaggg cgactgccct6720gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt6780gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga6840aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg6900agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc6960gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg7020aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg7080cattggcggc cttgctgttc ttctacggca agtgctgtgc acggatctgc cctggcttca7140ggagatcgga agacctcggc cgtccgggcg cttgccggtg gtgctgaccc cggatgaagt7200ctctagagct ctagagggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg7260cccagcttct gtatggaacg ggcatgcgga tcagtgaggg tttgcaactg cgggtcaagg7320atctggattt cgatcacggc acgatcatcg tgcgggaggg caagggctcc aaggatcggg7380ccttgatgtt acccgagagc ttggcaccca gcctgcgcga gcagggatcg atccaacccc7440tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcttct gaaaacgaca7500tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc7560ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg7620ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc7680aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg7740agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac7800gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc7860tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc7920cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg7980ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg8040cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc8100gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc8160atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca8220ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg8280agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt8340tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc8400cgcgcacgtc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct8460ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg8520atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt8580aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc8640
gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc 8700gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg 8760gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag 8820gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct 8880gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac 8940atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga 9000aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag 9060gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc 9120gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc 9180gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt 9240aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg 9300cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg 9360tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag 9420gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca 9480aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa 9540caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc 9600gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa 9660tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga 9720cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc 9780acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc 9840gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt 9900tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc 9960cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc10020agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga10080cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa10140gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg10200gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac10260cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc10320cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga10380gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc10440ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct10500ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat10560ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct10620gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc10680gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg10740tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt10800tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta10860cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa10920agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac10980ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc11040tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc11100tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc11160gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg11220atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag11280cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg11340gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc11400atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt11460aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc11520ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac11580agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa11640ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca11700caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc11760
gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata11820cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta11880tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca11940gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga12000cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg12060tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg12120tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg12180caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag12240aaaaaaagga tctcaagaag atccggaaaa cgcaagcgca aagagaaagc aggtagcttg12300cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga12360attgcc 12366<210>30<211>12384<212>DNA<213>人工<220>
<223>pTVE497<400>30agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg 120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga 180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg 240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat 300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt 360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat 420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag 480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agctcattct 540ttgtttccaa gaacttgctt aacaggttcg gtttggtcca catatagtct atgttcttgg 600atatacctga tgaccgaatt aggtaacaaa tattgtacag acatggcgcg tctgataaat 660agacgaactt tcgtggaaga aatatcatta tagatgagtt gcttgatgat aagaatattc 720cttctatgtt catacataat atcatgggat aacaaaaaag accttacatc agaaccagta 780cgttcgacaa tcaaacaacc gtaattaccg agaatgtgat gtaaatcggc gtccgcccaa 840acgtttggtt cacccattga ctctattagg tcaccaccag ccagcaacat tattttcaca 900ccaatttttt ctccagtaac agtagctaca ccacctctct taatattgat ttcgtgattg 960aaatgatcca agaccttggc agttcttgtg tatgaaggtt gcaatgactc ccatgcatcc1020accatcaacc aagatgaggt tctttcgcag gccaattcac acatacgtac tctatggtag1080gatggggcca agccttgctt ttgatagtta tcactaacag gggagtaata tccacctatg1140acttcaaacc ttgtttgttc agagattgca tctaaagcca tttcaaacat tcttagatgc1200aagtaggtga ttggtgaaaa agacccacat gctactatta ctaacggcag tttatttgga1260tccagtaatt ttttcgataa tctgtgtgag gggaattcgt agtcttccaa ggttcttgct1320tgacgaacga ttccatgtgg aacttcttct aaatcagcaa tctgatattt ctgaacacct1380ctagtttctg attctgtggt tgaatctgaa ccgtatctct ccttggatct aaaatccgcg1440tcatcatctt cggaagacac ttctgcggaa agtggttgaa agtcgtcttg gttcaatgga1500atatgctgat gttttttatc attgccttct ttacgactgt gatgatggtg atgatgatgc1560ttaggatgct ttttcttcct cttaatatta aaaggtgcat ctatagagga attcgcatcg1620gctaagacgt atggaataat tggaatagat ttgggaattt tagattccgg gtcgggtgga1680ggaatcaatt cctcgtctgc agatggcggt ttgaaatccg gagctcttgt aaccttgcgc1740ttcttcttgg gatccatggt tttggtttaa taagaagaga aaagagttct tttgttatgg1800
ctgaagtaat agagaaatga gctcgagtcc tctccaaatg aaatgaactt ccttatatag1860aggaagggtc ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatat1920cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc1980tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct2040ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga2100tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt2160gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga2220cgagagtgtc gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaaa2280gactctgtat gaactgttcg ccagtcttca cggcgagttc tgttagatcc tcgatctgaa2340tttttgactc catgtatggt gcatatggcg cgccatatgc ccgggccctg tacagcggcc2400gcgttaacgc gtatactcta gagcgatcgc ccgggccggc catttaaatg aattcgagct2460cggtacccaa acgcggccgc aagctataac ttcgtatagc atacattata cgaagttatt2520cgactctaga ggatcccaat tcccatgcat ggagtcaaag attcaaatag aggacacttc2580tcgaactcgg ccgtcgaact cggccgtcga gtacatggtc gataagaaaa ggcaatttgt2640agatgttaat tcccatcttg aaagaaatat agtttaaata tttattgata aaataacaag2700tcaggtatta tagtccaagc aaaaacataa atttattgat gcaagtttaa attcagaaat2760atttcaataa ctgattatat cagctggtac attgccgtag atgaaagact gagtgcgata2820ttatgtgtaa tacataaatt gatgatatag ctagcttagc tcatcggggg atcctagacg2880cgtgagatca gatctcggtg acgggcagga ccggacgggg cggtaccggc aggctgaagt2940ccagctgcca gaaacccacg tcatgccagt tcccgtgctt gaagccggcc gcccgcagca3000tgccgcgggg ggcatatccg agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc3060cgatgacagc gaccacgctc ttgaagccct gtgcctccag ggacttcagc aggtgggtgt3120agagcgtgga gcccagtccc gtccgctggt ggcgggggga gacgtacacg gtcgactcgg3180ccgtccagtc gtaggcgttg cgtgccttcc aggggcccgc gtaggcgatg ccggcgacct3240cgccgtccac ctcggcgacg agccagggat agcgctcccg cagacggacg aggtcgtccg3300tccactcctg cggttcctgc ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt3360ggttgacgat ggtgcagacc gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg3420ggcgtcgttc tgggtccatt gttcttcttt actctttgtg tgactgaggt ttggtctagt3480gctttggtca tctatatata atgataacaa caatgagaac aagctttgga gtgatcggag3540ggtctaggat acatgagatt caagtggact aggatctaca ccgttggatt ttgagtgtgg3600atatgtgtga ggttaatttt acttggtaac ggccacaaag gcctaaggag aggtgttgag3660acccttatcg gcttgaaccg ctggaataat gccacgtgga agataattcc atgaatctta3720tcgttatcta tgagtgaaat tgtgtgatgg tggagtggtg cttgctcatt ttacttgcct3780ggtggacttg gccctttcct tatggggaat ttatatttta cttactatag agctttcata3840cctttttttt accttggatt tagttaatat ataatggtat gattcatgaa taaaaatggg3900aaatttttga atttgtactg ctaaatgcat aagattaggt gaaactgtgg aatatatatt3960tttttcattt aaaagcaaaa tttgcctttt actagaatta taaatataga aaaatatata4020acattcaaat aaaaatgaaa ataagaactt tcaaaaaaca gaactatgtt taatgtgtaa4080agattagtcg cacatcaagt catctgttac aatatgttac aacaagtcat aagcccaaca4140aagttagcac gtctaaataa actaaagagt ccacgaaaat attacaaatc ataagcccaa4200caaagttatt gatcaaaaaa aaaaaacgcc caacaaagct aaacaaagtc caaaaaaaac4260ttctcaagtc tccatcttcc tttatgaaca ttgaaaacta tacacaaaac aagtcagata4320aatctctttc tgggcctgtc ttcccaacct cctacatcac ttccctatcg gattgaatgt4380tttacttgta ccttttccgt tgcaatgata ttgatagtat gtttgtgaaa actaataggg4440ttaacaatcg aagtcatgga atatggattt ggtccaagat tttccgagag ctttctagta4500gaaagcccat caccagaaat ttactagtaa aataaatcac caattaggtt tcttattatg4560tgccaaattc aatataatta tagaggatat ttcaaatgaa aacgtatgaa tgttattagt4620aaatggtcag gtaagacatt aaaaaaatcc tacgtcagat attcaacttt aaaaattcga4680tcagtgtgga attgtacaaa aatttgggat ctactatata tatataatgc tttacaacac4740ttggattttt ttttggaggc tggaattttt aatctacata tttgttttgg ccatgcacca4800actcattgtt tagtgtaata ctttgatttt gtcaaatata tgtgttcgtg tatatttgta4860taagaatttc tttgaccata tacacacaca catatatata tatatatata tattatatat4920
catgcacttt taattgaaaa aataatatat atatatatag tgcatttttt ctaacaacca4980tatatgttgc gattgatctg caaaaatact gctagagtaa tgaaaaatat aatctattgc5040tgaaattatc tcagatgtta agattttctt aaagtaaatt ctttcaaatt ttagctaaaa5100gtcttgtaat aactaaagaa taatacacaa tctcgaccac ggaaaaaaaa cacataataa5160atttgaattt cgaccgcggt acccggaatt gggttataat tacctcaggt cgaggaatta5220attcggtacg tacctaataa cttcgtatag catacattat acgaagttat atggatctcg5280aggcattacg gcattacggc actcgcgagg gtcccaattc gagcatggag ccatttacaa5340ttgaatatat cctgccgccg ctgccgcttt gcacccggtg gagcttgcat gttggtttct5400acgcagaact gagccggtta ggcagataat ttccattgag aactgagcca tgtgcacctt5460ccccccaaca cggtgagcga cggggcaacg gagtgatcca catgggactt ttaaacatca5520tccgtcggat ggcgttgcga gagaagcagt cgatccgtga gatcagccga cgcaccgggc5580aggcgcgcaa cacgatcgca aagtatttga acgcaggtac aatcgagccg acgttcacgg5640taccggaacg accaagcaag ctagcttagt aaagccctcg ctagatttta atgcggatgt5700tgcgattact tcgccaacta ttgcgataac aagaaaaagc cagcctttca tgatatatct5760cccaatttgt gtagggctta ttatgcacgc ttaaaaataa taaaagcaga cttgacctga5820tagtttggct gtgagcaatt atgtgcttag tgcatctaac gcttgagtta agccgcgccg5880cgaagcggcg tcggcttgaa cgaattgtta gacattattt gccgactacc ttggtgatct5940cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt6000cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca6060ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc6120tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg6180gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg6240gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg6300tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt6360cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga6420tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg6480aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta6540cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg6600agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta6660cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatgatg tttaactttg6720ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac6780ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac6840agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg6900ttctggacca gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg6960cttatgtcca ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct7020tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg7080tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag tgctgtgcac7140ggatctgccc tggcttcagg agatcggaag acctcggccg tccgggcgct tgccggtggt7200gctgaccccg gatgaagtct ctagagctct agagggttcg catcctcggt tttctggaag7260gcgagcatcg tttgttcgcc cagcttctgt atggaacggg catgcggatc agtgagggtt7320tgcaactgcg ggtcaaggat ctggatttcg atcacggcac gatcatcgtg cgggagggca7380agggctccaa ggatcgggcc ttgatgttac ccgagagctt ggcacccagc ctgcgcgagc7440agggatcgat ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc7500cgtcttctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc7560ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact7620agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc7680gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct7740gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca7800ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg7860cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc7920tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga7980ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg8040
ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg 8100cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg 8160ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag 8220tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg 8280acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga 8340cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg 8400gtacgtgttc gagccgcccg cgcacgtctc aaccgtgcgg ctgcatgaaa tcctggccgg 8460tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg 8520ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc 8580gtatatgatg cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct 8640gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc 8700ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc 8760gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg 8820attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc 8880caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg 8940cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc 9000attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc 9060acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag 9120tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt 9180gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa 9240tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa 9300gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca 9360cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga 9420tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc 9480taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc 9540ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga 9600acgggcggtt ggccaggcgt aagcggctgg gttgtctgcc ggccctgcaa tggcactgga 9660acccccaagc ccgaggaatc ggcgtgacgg tcgcaaacca tccggcccgg tacaaatcgg 9720cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg cccagcggca 9780acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg atcgaatccg 9840caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg 9900cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc gcgatagtcg 9960cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt10020gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg ccggcatggc10080cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg aatccatgaa10140ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac acgttgcgga10200cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc tggtagaaac10260ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg10320ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga tcgtaaagag10380cgaaaccggg cggccggagt acatcgagat cgagctagct gattggatgt accgcgagat10440cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt tgatcgatcc10500cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg cagaagccag10560atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca agaagttctg10620tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga10680ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg agggcgaagc10740atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag caggggaaaa10800aggtcgaaaa ggtctctttc ctgtggatag cacgtacatt gggaacccaa agccgtacat10860tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc ggtcacacat10920gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact ctttaaaact10980tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc gcacagccga11040agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg11100tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag gcaatctacc11160
agggcgcgga caagccgcgc cgtcgccact cgaccgccgg cgcccacatc aaggcaccct11220gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg11280tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg11340gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata11400ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga11460aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct11520cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc11580ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg11640ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg11700cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg11760actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac11820cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca11880tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt11940gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc12000caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag12060agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac12120tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt12180tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa12240gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat ccggaaaacg caagcgcaaa12300gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg12360gacagcaagc gaaccggaat tgcc 12384<210>31<211>12348<212>DNA<213>人工<220>
<223>pTVE 500<400>31agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg 120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga 180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg 240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat 300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt 360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat 420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag 480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agctcactct 540ttgctatcca agacctgctt gaccggttca ctttgattaa tgtatagatt atactcttgg 600atgtaacgga tgacagagtt tggaagaaga tattgaactg acattccacg tctgatgaaa 660agccgcactt tcgtagagga aatatcattg taaataagtt gtttgataat aaggatattt 720cttctgtgtt catacatgat atcatgggaa agcgagaagg acctaacatc agaaccagtc 780ctttccacga tcaaacatcc ataattaccc aaaatatggt gcaggtctga atcagcccac 840acatgaggct cgcccatgga ttcgataaga tcaccgcctg ccaataacat gatttttacg 900cccatttttt caccatctac agtcatgatt ccacctctct tgatatttat ttcatgattg 960aaatggtcca agacttttgc tgtccttgta taacttgatt gtaaagattc ccaggcatca1020accattaacc aagatgatgt ccgctcgcat gctaattcgc acatgcggac acgatgataa1080gctggggcta accctcgctt ttgatagtta tcacttactg gagaaaaata accaccaacc1140acttcaaaac gcgtttgctc attgatatca tctaaagcca tttcaaacat tctcaaatgt1200
aggtatgtta tgggagaaaa tgatccacaa gcaacgatga tcagaggcag tttttcagga1260tcttgtaact ttttcgtcaa tctgtgtaca ggaaattcgt aatcttcaat agttctggct1320tgtcttacaa ttgtgtgagg aacttcttcc aaatcagcaa tttggctctt cagtaccccc1380aaattcccag tcatcgtaac atcctgtaga gtagcgtcaa ttccattcga ttgcccttca1440ctttcctcct ccgatgatac atcccgagat agtggctgga agtcactagt atttaatgga1500atatggtcca ttcggttgct tgagttggcg ttttttttcc ttgacgacag ggttttgtaa1560atgtcgagat tgaaaggagc atcgatcgaa gaattataat cagctaaaac atatggaact1620atgggtccag attttggcat cgtatgtgtt ggatctggcg gtggttgtag ttcttcattt1680ggctgtggcg gtttaaaatc gggtgctttg gtgggatcca tggttttggt ttaataagaa1740gagaaaagag ttcttttgtt atggctgaag taatagagaa atgagctcga gtcctctcca1800aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg gattgtgcgt1860catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac gtggttggaa1920cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg1980cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg taggtgccac2040cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg aatccgagga2100ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct tctgagactg2160tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgttg acgaagattt2220tcttcttgtc attgagtcgt aaaagactct gtatgaactg ttcgccagtc ttcacggcga2280gttctgttag atcctcgatc tgaatttttg actccatgta tggtgcatat ggcgcgccat2340atgcccgggc cctgtacagc ggccgcgtta acgcgtatac tctagagcga tcgcccgggc2400cggccattta aatgaattcg agctcggtac ccaaacgcgg ccgcaagcta taacttcgta2460tagcatacat tatacgaagt tattcgactc tagaggatcc caattcccat gcatggagtc2520aaagattcaa atagaggaca cttctcgaac tcggccgtcg aactcggccg tcgagtacat2580ggtcgataag aaaaggcaat ttgtagatgt taattcccat cttgaaagaa atatagttta2640aatatttatt gataaaataa caagtcaggt attatagtcc aagcaaaaac ataaatttat2700tgatgcaagt ttaaattcag aaatatttca ataactgatt atatcagctg gtacattgcc2760gtagatgaaa gactgagtgc gatattatgt gtaatacata aattgatgat atagctagct2820tagctcatcg ggggatccta gacgcgtgag atcagatctc ggtgacgggc aggaccggac2880ggggcggtac cggcaggctg aagtccagct gccagaaacc cacgtcatgc cagttcccgt2940gcttgaagcc ggccgcccgc agcatgccgc ggggggcata tccgagcgcc tcgtgcatgc3000gcacgctcgg gtcgttgggc agcccgatga cagcgaccac gctcttgaag ccctgtgcct3060ccagggactt cagcaggtgg gtgtagagcg tggagcccag tcccgtccgc tggtggcggg3120gggagacgta cacggtcgac tcggccgtcc agtcgtaggc gttgcgtgcc ttccaggggc3180ccgcgtaggc gatgccggcg acctcgccgt ccacctcggc gacgagccag ggatagcgct3240cccgcagacg gacgaggtcg tccgtccact cctgcggttc ctgcggctcg gtacggaagt3300tgaccgtgct tgtctcgatg tagtggttga cgatggtgca gaccgccggc atgtccgcct3360cggtggcacg gcggatgtcg gccgggcgtc gttctgggtc cattgttctt ctttactctt3420tgtgtgactg aggtttggtc tagtgctttg gtcatctata tataatgata acaacaatga3480gaacaagctt tggagtgatc ggagggtcta ggatacatga gattcaagtg gactaggatc3540tacaccgttg gattttgagt gtggatatgt gtgaggttaa ttttacttgg taacggccac3600aaaggcctaa ggagaggtgt tgagaccctt atcggcttga accgctggaa taatgccacg3660tggaagataa ttccatgaat cttatcgtta tctatgagtg aaattgtgtg atggtggagt3720ggtgcttgct cattttactt gcctggtgga cttggccctt tccttatggg gaatttatat3780tttacttact atagagcttt catacctttt ttttaccttg gatttagtta atatataatg3840gtatgattca tgaataaaaa tgggaaattt ttgaatttgt actgctaaat gcataagatt3900aggtgaaact gtggaatata tatttttttc atttaaaagc aaaatttgcc ttttactaga3960attataaata tagaaaaata tataacattc aaataaaaat gaaaataaga actttcaaaa4020aacagaacta tgtttaatgt gtaaagatta gtcgcacatc aagtcatctg ttacaatatg4080ttacaacaag tcataagccc aacaaagtta gcacgtctaa ataaactaaa gagtccacga4140aaatattaca aatcataagc ccaacaaagt tattgatcaa aaaaaaaaaa cgcccaacaa4200agctaaacaa agtccaaaaa aaacttctca agtctccatc ttcctttatg aacattgaaa4260actatacaca aaacaagtca gataaatctc tttctgggcc tgtcttccca acctcctaca4320
tcacttccct atcggattga atgttttact tgtacctttt ccgttgcaat gatattgata4380gtatgtttgt gaaaactaat agggttaaca atcgaagtca tggaatatgg atttggtcca4440agattttccg agagctttct agtagaaagc ccatcaccag aaatttacta gtaaaataaa4500tcaccaatta ggtttcttat tatgtgccaa attcaatata attatagagg atatttcaaa4560tgaaaacgta tgaatgttat tagtaaatgg tcaggtaaga cattaaaaaa atcctacgtc4620agatattcaa ctttaaaaat tcgatcagtg tggaattgta caaaaatttg ggatctacta4680tatatatata atgctttaca acacttggat ttttttttgg aggctggaat ttttaatcta4740catatttgtt ttggccatgc accaactcat tgtttagtgt aatactttga ttttgtcaaa4800tatatgtgtt cgtgtatatt tgtataagaa tttctttgac catatacaca cacacatata4860tatatatata tatatattat atatcatgca cttttaattg aaaaaataat atatatatat4920atagtgcatt ttttctaaca accatatatg ttgcgattga tctgcaaaaa tactgctaga4980gtaatgaaaa atataatcta ttgctgaaat tatctcagat gttaagattt tcttaaagta5040aattctttca aattttagct aaaagtcttg taataactaa agaataatac acaatctcga5100ccacggaaaa aaaacacata ataaatttga atttcgaccg cggtacccgg aattgggtta5160taattacctc aggtcgagga attaattcgg tacgtaccta ataacttcgt atagcataca5220ttatacgaag ttatatggat ctcgaggcat tacggcatta cggcactcgc gagggtccca5280attcgagcat ggagccattt acaattgaat atatcctgcc gccgctgccg ctttgcaccc5340ggtggagctt gcatgttggt ttctacgcag aactgagccg gttaggcaga taatttccat5400tgagaactga gccatgtgca ccttcccccc aacacggtga gcgacggggc aacggagtga5460tccacatggg acttttaaac atcatccgtc ggatggcgtt gcgagagaag cagtcgatcc5520gtgagatcag ccgacgcacc gggcaggcgc gcaacacgat cgcaaagtat ttgaacgcag5580gtacaatcga gccgacgttc acggtaccgg aacgaccaag caagctagct tagtaaagcc5640ctcgctagat tttaatgcgg atgttgcgat tacttcgcca actattgcga taacaagaaa5700aagccagcct ttcatgatat atctcccaat ttgtgtaggg cttattatgc acgcttaaaa5760ataataaaag cagacttgac ctgatagttt ggctgtgagc aattatgtgc ttagtgcatc5820taacgcttga gttaagccgc gccgcgaagc ggcgtcggct tgaacgaatt gttagacatt5880atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat5940ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta6000tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg6060gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc6120gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct6180caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg6240caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg6300gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct6360tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc6420ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc6480gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg6540gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga6600gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg6660cttccctcat gatgtttaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc6720tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag6780gcatagactg taccccaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt6840taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc6900attacagctt acgaaccgaa caggcttatg tccactgggt tcgtgccttc atccgtttcc6960acggtgtgcg tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc7020tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt7080tcttctacgg caagtgctgt gcacggatct gccctggctt caggagatcg gaagacctcg7140gccgtccggg cgcttgccgg tggtgctgac cccggatgaa gtctctagag ctctagaggg7200ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccagctt ctgtatggaa7260cgggcatgcg gatcagtgag ggtttgcaac tgcgggtcaa ggatctggat ttcgatcacg7320gcacgatcat cgtgcgggag ggcaagggct ccaaggatcg ggccttgatg ttacccgaga7380gcttggcacc cagcctgcgc gagcagggat cgatccaacc cctccgctgc tatagtgcag7440
tcggcttctg acgttcagtg cagccgtctt ctgaaaacga catgtcgcac aagtcctaag 7500ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc 7560gcataaagta gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg 7620ccgctggcct gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac 7680gggccgaact gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca 7740ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga 7800cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc 7860gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca 7920cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc 7980taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg 8040gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg 8100aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc 8160gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc 8220gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg 8280aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat 8340cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt 8400gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag 8460cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa 8520cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa 8580ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc 8640atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat 8700tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc 8760gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac 8820ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca 8880gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac 8940ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt 9000gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc 9060gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact 9120gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc 9180caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa 9240tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg 9300caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg 9360cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc 9420tgctatctga atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga 9480tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc 9540gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgtc 9600tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg acggtcgcaa 9660accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa 9720ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg 9780gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc 9840gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta 9900tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa 9960gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt10020ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt10080ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg10140cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa10200gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca10260gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat10320tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct10380agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca10440ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg10500cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag10560
cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct10620gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta10680ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg10740gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctc tttcctgtgg atagcacgta10800cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc10860gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt10920ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata10980actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg11040ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc11100tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg11160ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct11220gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac11280aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt11340cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact11400gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat11460caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg11520agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc11580aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt11640gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag11700tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc11760cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc11820ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt11880cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt11940atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc12000agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa12060gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa12120gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg12180tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga12240agatccggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg12300atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgcc 12348<210>32<211>12366<212>DNA<213>人工<220>
<223>pTVE 501<400>32agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agctcactct540ttgctatcca agacctgctt gaccggttca ctttgattaa tgtatagatt atactcttgg600atgtaacgga tgacagagtt tggaagaaga tattgaactg acattccacg tctgatgaaa660
agccgcactt tcgtagagga aatatcattg taaataagtt gtttgataat aaggatattt 720cttctgtgtt catacatgat atcatgggaa agcgagaagg acctaacatc agaaccagtc 780ctttccacga tcaaacatcc ataattaccc aaaatatggt gcaggtctga atcagcccac 840acatgaggct cgcccatgga ttcgataaga tcaccgcctg ccaataacat gatttttacg 900cccatttttt caccatctac agtcatgatt ccacctctct tgatatttat ttcatgattg 960aaatggtcca agacttttgc tgtccttgta taacttgatt gtaaagattc ccaggcatca1020accattaacc aagatgatgt ccgctcgcat gctaattcgc acatgcggac acgatgataa1080gctggggcta accctcgctt ttgatagtta tcacttactg gagaaaaata accaccaacc1140acttcaaaac gcgtttgctc attgatatca tctaaagcca tttcaaacat tctcaaatgt1200aggtatgtta tgggagaaaa tgatccacaa gcaacgatga tcagaggcag tttttcagga1260tcttgtaact ttttcgtcaa tctgtgtaca ggaaattcgt aatcttcaat agttctggct1320tgtcttacaa ttgtgtgagg aacttcttcc aaatcagcaa tttggctctt cagtaccccc1380aaattcccag tcatcgtaac atcctgtaga gtagcgtcaa ttccattcga ttgcccttca1440ctttcctcct ccgatgatac atcccgagat agtggctgga agtcactagt atttaatgga1500atatggtcca ttcggttgct tgagttggcg ttttttttcc ttgacgacag ggttttgtaa1560atgtcgagat tgaaaggagc atcgatcgaa gaattataat cagctaaaac atatggaact1620atgggtccag attttggcat cgtatgtgtt ggatctggcg gtggttgtag ttcttcattt1680ggctgtggcg gtttaaaatc gggtgctttg gtaaccttgc gcttcttctt gggatccatg1740gttttggttt aataagaaga gaaaagagtt cttttgttat ggctgaagta atagagaaat1800gagctcgagt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag1860gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct1920ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat1980ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat2040ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg2100ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc2160tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg tcgtgctcca2220ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt2280cgccagtctt cacggcgagt tctgttagat cctcgatctg aatttttgac tccatgtatg2340gtgcatatgg cgcgccatat gcccgggccc tgtacagcgg ccgcgttaac gcgtatactc2400tagagcgatc gcccgggccg gccatttaaa tgaattcgag ctcggtaccc aaacgcggcc2460gcaagctata acttcgtata gcatacatta tacgaagtta ttcgactcta gaggatccca2520attcccatgc atggagtcaa agattcaaat agaggacact tctcgaactc ggccgtcgaa2580ctcggccgtc gagtacatgg tcgataagaa aaggcaattt gtagatgtta attcccatct2640tgaaagaaat atagtttaaa tatttattga taaaataaca agtcaggtat tatagtccaa2700gcaaaaacat aaatttattg atgcaagttt aaattcagaa atatttcaat aactgattat2760atcagctggt acattgccgt agatgaaaga ctgagtgcga tattatgtgt aatacataaa2820ttgatgatat agctagctta gctcatcggg ggatcctaga cgcgtgagat cagatctcgg2880tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca2940cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc3000cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc3060tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc3120ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt3180tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga3240cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct3300gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga3360ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggtcca3420ttgttcttct ttactctttg tgtgactgag gtttggtcta gtgctttggt catctatata3480taatgataac aacaatgaga acaagctttg gagtgatcgg agggtctagg atacatgaga3540ttcaagtgga ctaggatcta caccgttgga ttttgagtgt ggatatgtgt gaggttaatt3600ttacttggta acggccacaa aggcctaagg agaggtgttg agacccttat cggcttgaac3660cgctggaata atgccacgtg gaagataatt ccatgaatct tatcgttatc tatgagtgaa3720attgtgtgat ggtggagtgg tgcttgctca ttttacttgc ctggtggact tggccctttc3780
cttatgggga atttatattt tacttactat agagctttca tacctttttt ttaccttgga3840tttagttaat atataatggt atgattcatg aataaaaatg ggaaattttt gaatttgtac3900tgctaaatgc ataagattag gtgaaactgt ggaatatata tttttttcat ttaaaagcaa3960aatttgcctt ttactagaat tataaatata gaaaaatata taacattcaa ataaaaatga4020aaataagaac tttcaaaaaa cagaactatg tttaatgtgt aaagattagt cgcacatcaa4080gtcatctgtt acaatatgtt acaacaagtc ataagcccaa caaagttagc acgtctaaat4140aaactaaaga gtccacgaaa atattacaaa tcataagccc aacaaagtta ttgatcaaaa4200aaaaaaaacg cccaacaaag ctaaacaaag tccaaaaaaa acttctcaag tctccatctt4260cctttatgaa cattgaaaac tatacacaaa acaagtcaga taaatctctt tctgggcctg4320tcttcccaac ctcctacatc acttccctat cggattgaat gttttacttg taccttttcc4380gttgcaatga tattgatagt atgtttgtga aaactaatag ggttaacaat cgaagtcatg4440gaatatggat ttggtccaag attttccgag agctttctag tagaaagccc atcaccagaa4500atttactagt aaaataaatc accaattagg tttcttatta tgtgccaaat tcaatataat4560tatagaggat atttcaaatg aaaacgtatg aatgttatta gtaaatggtc aggtaagaca4620ttaaaaaaat cctacgtcag atattcaact ttaaaaattc gatcagtgtg gaattgtaca4680aaaatttggg atctactata tatatataat gctttacaac acttggattt ttttttggag4740gctggaattt ttaatctaca tatttgtttt ggccatgcac caactcattg tttagtgtaa4800tactttgatt ttgtcaaata tatgtgttcg tgtatatttg tataagaatt tctttgacca4860tatacacaca cacatatata tatatatata tatattatat atcatgcact tttaattgaa4920aaaataatat atatatatat agtgcatttt ttctaacaac catatatgtt gcgattgatc4980tgcaaaaata ctgctagagt aatgaaaaat ataatctatt gctgaaatta tctcagatgt5040taagattttc ttaaagtaaa ttctttcaaa ttttagctaa aagtcttgta ataactaaag5100aataatacac aatctcgacc acggaaaaaa aacacataat aaatttgaat ttcgaccgcg5160gtacccggaa ttgggttata attacctcag gtcgaggaat taattcggta cgtacctaat5220aacttcgtat agcatacatt atacgaagtt atatggatct cgaggcatta cggcattacg5280gcactcgcga gggtcccaat tcgagcatgg agccatttac aattgaatat atcctgccgc5340cgctgccgct ttgcacccgg tggagcttgc atgttggttt ctacgcagaa ctgagccggt5400taggcagata atttccattg agaactgagc catgtgcacc ttccccccaa cacggtgagc5460gacggggcaa cggagtgatc cacatgggac ttttaaacat catccgtcgg atggcgttgc5520gagagaagca gtcgatccgt gagatcagcc gacgcaccgg gcaggcgcgc aacacgatcg5580caaagtattt gaacgcaggt acaatcgagc cgacgttcac ggtaccggaa cgaccaagca5640agctagctta gtaaagccct cgctagattt taatgcggat gttgcgatta cttcgccaac5700tattgcgata acaagaaaaa gccagccttt catgatatat ctcccaattt gtgtagggct5760tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa5820ttatgtgctt agtgcatcta acgcttgagt taagccgcgc cgcgaagcgg cgtcggcttg5880aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga5940caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag6000cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt6060cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca6120acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta6180aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg6240ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat6300caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc6360caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa6420tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa6480ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca6540aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg6600ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg6660atacttcggc gatcaccgct tccctcatga tgtttaactt tgttttaggg cgactgccct6720gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt6780gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga6840aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg6900
agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc6960gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg7020aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg7080cattggcggc cttgctgttc ttctacggca agtgctgtgc acggatctgc cctggcttca7140ggagatcgga agacctcggc cgtccgggcg cttgccggtg gtgctgaccc cggatgaagt7200ctctagagct ctagagggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg7260cccagcttct gtatggaacg ggcatgcgga tcagtgaggg tttgcaactg cgggtcaagg7320atctggattt cgatcacggc acgatcatcg tgcgggaggg caagggctcc aaggatcggg7380ccttgatgtt acccgagagc ttggcaccca gcctgcgcga gcagggatcg atccaacccc7440tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcttct gaaaacgaca7500tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc7560ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg7620ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc7680aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg7740agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac7800gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc7860tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc7920cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg7980ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg8040cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc8100gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc8160atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca8220ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg8280agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt8340tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc8400cgcgcacgtc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct8460ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg8520atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt8580aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc8640gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc8700gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg8760gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag8820gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct8880gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac8940atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga9000aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag9060gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc9120gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc9180gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt9240aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg9300cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg9360tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag9420gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca9480aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa9540caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc9600gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa9660tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga9720cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc9780acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc9840gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt9900tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc9960cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc 10020
agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga10080cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa10140gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg10200gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac10260cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc10320cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga10380gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc10440ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct10500ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat10560ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct10620gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc10680gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg10740tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt10800tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta10860cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa10920agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac10980ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc11040tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc11100tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc11160gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg11220atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag11280cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg11340gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc11400atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt11460aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc11520ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac11580agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa11640ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca11700caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc11760gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata11820cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta11880tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca11940gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga12000cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg12060tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg12120tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg12180caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag12240aaaaaaagga tctcaagaag atccggaaaa cgcaagcgca aagagaaagc aggtagcttg12300cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga12360attgcc 12366<210>33<211>13305<212>DNA<213>人工<220>
<223>pTVE502<400>33agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc60
cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg 120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga 180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg 240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat 300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt 360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat 420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag 480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agcctaatca 540atagacataa tgtcaagcgt tgagccttta tgtgcttcac actgctcgac aacttcatca 600atttttcttg aagcccatgg aaatcttggg ttgattaaga aaggacgtaa gtcaaatctg 660ttgtcttctg gtgaatactg ttcagcatga taactaggag ttaaaacagt ttgcttgtgt 720ctgttgatgg catagaagaa gaaaaatctt ttcacctttt cagatatttg acgaggtgtt 780aactttgggg accattgatg aagaagtttt aagaacatag aataaggacc acatttttca 840acctttctta ggtaaccaaa cacgcccaat tcttcatacg tcatccccat atctatctca 900tccgattgaa cgtaatcttt agtcataggt tctaattctg cagttggtgt agcgtttaaa 960aagtcattca agattggcat gttatattgt tttgatgcgt aggcaatgaa tcttttcaag1020tcagtttttg aaataccccc aatagggttg atatctgcgg aggagcagtc atattttgtt1080agatacccac gtaagcactc atcaacattt gcgctaccaa gtactaacaa tccacccgag1140tttgggatac cacgaaccca cggcaacagt tgcgcaaaaa gataagaaag aaccattctt1200agacgcgcct ggatgttttg taaagccaag ttctcgattt gagatccccc aaatattttg1260tatattggtt ttttgccagt ggctacttcg aataaggaca ccacactgga taccaatgag1320tccatcttta aatccacgtg gtaagatcca attgcattgg aaaggtcctt tgctctgttt1380cttgtctcct tggatgaatt ttccgtaccc atgaaacagg agtgaaatat ttttgaggct1440agatcctgtg gactgtctgg aatccaatca tcgccgctac gtgttatctt acgaacgtct1500ttgataactt gctcatttcc attttgagca gcgtcggtca ctaaacggca catagagtgg1560acaatcattg cagttgcaca agagtcaatg cccccagata agggaaggaa aaaccctgtt1620ccgttacaac gtcttaaata atcccacatc cagcaagcag gtcccagtgc aatttcttcc1680tcaggagagt gatagaaagg ctcgcggact tttgttggac acactgtagg atcaaatctg1740gaggtcatta aagccaattc tacaggaata tcaatacgct tgaactttat ttctgccaag1800gaggcttgta ggccacgaga catgacagct gcacgataac tcctcacctc ttctaggtcc1860acagtagcag taactacttc cacatcatct agcgaaaatt gtgaaccttg ggctacaatt1920gtaccattga tggcaattag tgcacagcca tcataatata atctgtcacc atcacaacct1980ctttgatttg catacaagta aacaccacca caacgtttag tggcatttaa aattaggtct2040aaccttttat ttaacttacg cagttcatga tgagaaccag atgagtttgt catgatttcc2100acaccatcta aagacatggc gatgtggggg gattgaggtg taaacaattc ttcacaagtt2160tctgtaccaa tgcatgtatc caatgaattt atcacagcgt ccccaaatgg cacaagtctc2220tggccggtaa ctttctgaat ctcaggtgga aggataaagt cctccaccac gccaggtttc2280atccaaggtg tgaaaaatct catttcccta tagttaccat cattagctaa ccaaatctta2340ggtcttatga acaatatctc accatccaag gataacaaac gacaattata acgaacattc2400ttgtgtagaa cgggcatacc aatgtcaagt attaatccat gggtttcttt attcttaatg2460atttgagcat acatttccca tgaatgaagg caaacgtcat tttctaaaaa atgatctaaa2520catccgtagc cagttatttc cagttctggg ccgacacgta acctggcacc cctctctttg2580gcaatcttaa tggactgtag gatacggtct ctattacctt caaaatctag ggcccattga2640ttcaagttgc atgtagctaa agtgataaga tgtgccatgg ttttggttta ataagaagag2700aaaagagttc ttttgttatg gctgaagtaa tagagaaatg agctcgagtc ctctccaaat2760gaaatgaact tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat2820cccttacgtc agtggagata tcacatcaat ccacttgctt tgaagacgtg gttggaacgt2880cttctttttc cacgatgctc ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag2940aggcatcttg aacgatagcc tttcctttat cgcaatgatg gcatttgtag gtgccacctt3000ccttttctac tgtccttttg atgaagtgac agatagctgg gcaatggaat ccgaggaggt3060ttcccgatat taccctttgt tgaaaagtct caatagccct ttggtcttct gagactgtat3120ctttgatatt cttggagtag acgagagtgt cgtgctccac catgttgacg aagattttct3180
tcttgtcatt gagtcgtaaa agactctgta tgaactgttc gccagtcttc acggcgagtt3240ctgttagatc ctcgatctga atttttgact ccatgtatgg tgcatatggc gcgccatatg3300cccgggccct gtacagcggc cgcgttaacg cgtatactct agagcgatcg cccgggccgg3360ccatttaaat gaattcgagc tcggtaccca aacgcggccg caagctataa cttcgtatag3420catacattat acgaagttat tcgactctag aggatcccaa ttcccatgca tggagtcaaa3480gattcaaata gaggacactt ctcgaactcg gccgtcgaac tcggccgtcg agtacatggt3540cgataagaaa aggcaatttg tagatgttaa ttcccatctt gaaagaaata tagtttaaat3600atttattgat aaaataacaa gtcaggtatt atagtccaag caaaaacata aatttattga3660tgcaagttta aattcagaaa tatttcaata actgattata tcagctggta cattgccgta3720gatgaaagac tgagtgcgat attatgtgta atacataaat tgatgatata gctagcttag3780ctcatcgggg gatcctagac gcgtgagatc agatctcggt gacgggcagg accggacggg3840gcggtaccgg caggctgaag tccagctgcc agaaacccac gtcatgccag ttcccgtgct3900tgaagccggc cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg tgcatgcgca3960cgctcgggtc gttgggcagc ccgatgacag cgaccacgct cttgaagccc tgtgcctcca4020gggacttcag caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg tggcgggggg4080agacgtacac ggtcgactcg gccgtccagt cgtaggcgtt gcgtgccttc caggggcccg4140cgtaggcgat gccggcgacc tcgccgtcca cctcggcgac gagccaggga tagcgctccc4200gcagacggac gaggtcgtcc gtccactcct gcggttcctg cggctcggta cggaagttga4260ccgtgcttgt ctcgatgtag tggttgacga tggtgcagac cgccggcatg tccgcctcgg4320tggcacggcg gatgtcggcc gggcgtcgtt ctgggtccat tgttcttctt tactctttgt4380gtgactgagg tttggtctag tgctttggtc atctatatat aatgataaca acaatgagaa4440caagctttgg agtgatcgga gggtctagga tacatgagat tcaagtggac taggatctac4500accgttggat tttgagtgtg gatatgtgtg aggttaattt tacttggtaa cggccacaaa4560ggcctaagga gaggtgttga gacccttatc ggcttgaacc gctggaataa tgccacgtgg4620aagataattc catgaatctt atcgttatct atgagtgaaa ttgtgtgatg gtggagtggt4680gcttgctcat tttacttgcc tggtggactt ggccctttcc ttatggggaa tttatatttt4740acttactata gagctttcat accttttttt taccttggat ttagttaata tataatggta4800tgattcatga ataaaaatgg gaaatttttg aatttgtact gctaaatgca taagattagg4860tgaaactgtg gaatatatat ttttttcatt taaaagcaaa atttgccttt tactagaatt4920ataaatatag aaaaatatat aacattcaaa taaaaatgaa aataagaact ttcaaaaaac4980agaactatgt ttaatgtgta aagattagtc gcacatcaag tcatctgtta caatatgtta5040caacaagtca taagcccaac aaagttagca cgtctaaata aactaaagag tccacgaaaa5100tattacaaat cataagccca acaaagttat tgatcaaaaa aaaaaaacgc ccaacaaagc5160taaacaaagt ccaaaaaaaa cttctcaagt ctccatcttc ctttatgaac attgaaaact5220atacacaaaa caagtcagat aaatctcttt ctgggcctgt cttcccaacc tcctacatca5280cttccctatc ggattgaatg ttttacttgt accttttccg ttgcaatgat attgatagta5340tgtttgtgaa aactaatagg gttaacaatc gaagtcatgg aatatggatt tggtccaaga5400ttttccgaga gctttctagt agaaagccca tcaccagaaa tttactagta aaataaatca5460ccaattaggt ttcttattat gtgccaaatt caatataatt atagaggata tttcaaatga5520aaacgtatga atgttattag taaatggtca ggtaagacat taaaaaaatc ctacgtcaga5580tattcaactt taaaaattcg atcagtgtgg aattgtacaa aaatttggga tctactatat5640atatataatg ctttacaaca cttggatttt tttttggagg ctggaatttt taatctacat5700atttgttttg gccatgcacc aactcattgt ttagtgtaat actttgattt tgtcaaatat5760atgtgttcgt gtatatttgt ataagaattt ctttgaccat atacacacac acatatatat5820atatatatat atattatata tcatgcactt ttaattgaaa aaataatata tatatatata5880gtgcattttt tctaacaacc atatatgttg cgattgatct gcaaaaatac tgctagagta5940atgaaaaata taatctattg ctgaaattat ctcagatgtt aagattttct taaagtaaat6000tctttcaaat tttagctaaa agtcttgtaa taactaaaga ataatacaca atctcgacca6060cggaaaaaaa acacataata aatttgaatt tcgaccgcgg tacccggaat tgggttataa6120ttacctcagg tcgaggaatt aattcggtac gtacctaata acttcgtata gcatacatta6180tacgaagtta tatggatctc gaggcattac ggcattacgg cactcgcgag ggtcccaatt6240cgagcatgga gccatttaca attgaatata tcctgccgcc gctgccgctt tgcacccggt6300
ggagcttgca tgttggtttc tacgcagaac tgagccggtt aggcagataa tttccattga6360gaactgagcc atgtgcacct tccccccaac acggtgagcg acggggcaac ggagtgatcc6420acatgggact tttaaacatc atccgtcgga tggcgttgcg agagaagcag tcgatccgtg6480agatcagccg acgcaccggg caggcgcgca acacgatcgc aaagtatttg aacgcaggta6540caatcgagcc gacgttcacg gtaccggaac gaccaagcaa gctagcttag taaagccctc6600gctagatttt aatgcggatg ttgcgattac ttcgccaact attgcgataa caagaaaaag6660ccagcctttc atgatatatc tcccaatttg tgtagggctt attatgcacg cttaaaaata6720ataaaagcag acttgacctg atagtttggc tgtgagcaat tatgtgctta gtgcatctaa6780cgcttgagtt aagccgcgcc gcgaagcggc gtcggcttga acgaattgtt agacattatt6840tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg6900cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga6960cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg7020cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct7080catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa7140atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa7200cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct7260cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag7320ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga7380gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc7440gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct7500tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat7560ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt7620ccctcatgat gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc7680tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca7740tagactgtac cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac7800caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt7860acagcttacg aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg7920gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg7980cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct8040tctacggcaa gtgctgtgca cggatctgcc ctggcttcag gagatcggaa gacctcggcc8100gtccgggcgc ttgccggtgg tgctgacccc ggatgaagtc tctagagctc tagagggttc8160gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccagcttctg tatggaacgg8220gcatgcggat cagtgagggt ttgcaactgc gggtcaagga tctggatttc gatcacggca8280cgatcatcgt gcgggagggc aagggctcca aggatcgggc cttgatgtta cccgagagct8340tggcacccag cctgcgcgag cagggatcga tccaacccct ccgctgctat agtgcagtcg8400gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta8460cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca8520taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg8580ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg8640ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc8700gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag8760tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca8820tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc8880cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa8940tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc9000cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag9060gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg9120cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg9180aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac9240aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga9300ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg9360gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt9420
ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag 9480cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg 9540gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc 9600gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc 9660gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt 9720gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc 9780gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc 9840gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg 9900gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc 9960gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg10020tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc10080gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag10140gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga10200gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa10260cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg10320aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc10380tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga10440attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg10500gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgtctgc10560cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgacg gtcgcaaacc10620atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc10680cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca10740agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc10800gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga10860cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg10920tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc10980cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc11040ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt11100gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca11160gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg11220tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag11280ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc11340tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc11400cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc11460cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc11520cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc11580ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg11640caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca11700aattgcccta gcaggggaaa aaggtcgaaa aggtctcttt cctgtggata gcacgtacat11760tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta11820cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc11880cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact11940gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc12000cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg12060cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg12120gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac12180acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag12240cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac12300gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag12360agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag12420gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc12480ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg12540
aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct12600ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca12660gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct12720cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc12780gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt12840tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc12900cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc12960cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg13020gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc13080agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag13140cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga13200tccggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata13260gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgcc13305<210>34<211>13326<212>DNA<213>人工<220>
<223>pTVE503<400>34agattcgaag ctcggtcccg tgggtgttct gtcgtctcgt tgtacaacga aatccattcc 60cattccgcgc tcaagatggc ttcccctcgg cagttcatca gggctaaatc aatctagccg 120acttgtccgg tgaaatgggc tgcactccaa cagaaacaat caaacaaaca tacacagcga 180cttattcaca cgcgacaaat tacaacggta tatatcctgc cagtactcgg ccgtcgaccg 240cggtaccccg gaattaagct tgcatgcctg caggcaattg gccgctgtac catgcatgat 300ctggatttta gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt 360tacaaataca aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat 420aggaacccta attcccttat ctgggaacta ctcacacatt attatggaga aaatagagag 480agatagattt gtagagagag actggtgatt tcagcgtgtc caagcttgct agcctaatca 540atagacataa tgtcaagcgt tgagccttta tgtgcttcac actgctcgac aacttcatca 600atttttcttg aagcccatgg aaatcttggg ttgattaaga aaggacgtaa gtcaaatctg 660ttgtcttctg gtgaatactg ttcagcatga taactaggag ttaaaacagt ttgcttgtgt 720ctgttgatgg catagaagaa gaaaaatctt ttcacctttt cagatatttg acgaggtgtt 780aactttgggg accattgatg aagaagtttt aagaacatag aataaggacc acatttttca 840acctttctta ggtaaccaaa cacgcccaat tcttcatacg tcatccccat atctatctca 900tccgattgaa cgtaatcttt agtcataggt tctaattctg cagttggtgt agcgtttaaa 960aagtcattca agattggcat gttatattgt tttgatgcgt aggcaatgaa tcttttcaag1020tcagtttttg aaataccccc aatagggttg atatctgcgg aggagcagtc atattttgtt1080agatacccac gtaagcactc atcaacattt gcgctaccaa gtactaacaa tccacccgag1140tttgggatac cacgaaccca cggcaacagt tgcgcaaaaa gataagaaag aaccattctt1200agacgcgcct ggatgttttg taaagccaag ttctcgattt gagatccccc aaatattttg1260tatattggtt ttttgccagt ggctacttcg aataaggaca ccacactgga taccaatgag1320tccatcttta aatccacgtg gtaagatcca attgcattgg aaaggtcctt tgctctgttt1380cttgtctcct tggatgaatt ttccgtaccc atgaaacagg agtgaaatat ttttgaggct1440agatcctgtg gactgtctgg aatccaatca tcgccgctac gtgttatctt acgaacgtct1500ttgataactt gctcatttcc attttgagca gcgtcggtca ctaaacggca catagagtgg1560acaatcattg cagttgcaca agagtcaatg cccccagata agggaaggaa aaaccctgtt1620ccgttacaac gtcttaaata atcccacatc cagcaagcag gtcccagtgc aatttcttcc1680
tcaggagagt gatagaaagg ctcgcggact tttgttggac acactgtagg atcaaatctg1740gaggtcatta aagccaattc tacaggaata tcaatacgct tgaactttat ttctgccaag1800gaggcttgta ggccacgaga catgacagct gcacgataac tcctcacctc ttctaggtcc1860acagtagcag taactacttc cacatcatct agcgaaaatt gtgaaccttg ggctacaatt1920gtaccattga tggcaattag tgcacagcca tcataatata atctgtcacc atcacaacct1980ctttgatttg catacaagta aacaccacca caacgtttag tggcatttaa aattaggtct2040aaccttttat ttaacttacg cagttcatga tgagaaccag atgagtttgt catgatttcc2100acaccatcta aagacatggc gatgtggggg gattgaggtg taaacaattc ttcacaagtt2160tctgtaccaa tgcatgtatc caatgaattt atcacagcgt ccccaaatgg cacaagtctc2220tggccggtaa ctttctgaat ctcaggtgga aggataaagt cctccaccac gccaggtttc2280atccaaggtg tgaaaaatct catttcccta tagttaccat cattagctaa ccaaatctta2340ggtcttatga acaatatctc accatccaag gataacaaac gacaattata acgaacattc2400ttgtgtagaa cgggcatacc aatgtcaagt attaatccat gggtttcttt attcttaatg2460atttgagcat acatttccca tgaatgaagg caaacgtcat tttctaaaaa atgatctaaa2520catccgtagc cagttatttc cagttctggg ccgacacgta acctggcacc cctctctttg2580gcaatcttaa tggactgtag gatacggtct ctattacctt caaaatctag ggcccattga2640ttcaagttgc atgtagctaa agtgataaga accttgcgct tcttcttggg atgtgccatg2700gttttggttt aataagaaga gaaaagagtt cttttgttat ggctgaagta atagagaaat2760gagctcgagt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag2820gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct2880ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat2940ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat3000ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg3060ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc3120tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg tcgtgctcca3180ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt3240cgccagtctt cacggcgagt tctgttagat cctcgatctg aatttttgac tccatgtatg3300gtgcatatgg cgcgccatat gcccgggccc tgtacagcgg ccgcgttaac gcgtatactc3360tagagcgatc gcccgggccg gccatttaaa tgaattcgag ctcggtaccc aaacgcggcc3420gcaagctata acttcgtata gcatacatta tacgaagtta ttcgactcta gaggatccca3480attcccatgc atggagtcaa agattcaaat agaggacact tctcgaactc ggccgtcgaa3540ctcggccgtc gagtacatgg tcgataagaa aaggcaattt gtagatgtta attcccatct3600tgaaagaaat atagtttaaa tatttattga taaaataaca agtcaggtat tatagtccaa3660gcaaaaacat aaatttattg atgcaagttt aaattcagaa atatttcaat aactgattat3720atcagctggt acattgccgt agatgaaaga ctgagtgcga tattatgtgt aatacataaa3780ttgatgatat agctagctta gctcatcggg ggatcctaga cgcgtgagat cagatctcgg3840tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca3900cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc3960cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc4020tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc4080ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt4140tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga4200cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct4260gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga4320ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggtcca4380ttgttcttct ttactctttg tgtgactgag gtttggtcta gtgctttggt catctatata4440taatgataac aacaatgaga acaagctttg gagtgatcgg agggtctagg atacatgaga4500ttcaagtgga ctaggatcta caccgttgga ttttgagtgt ggatatgtgt gaggttaatt4560ttacttggta acggccacaa aggcctaagg agaggtgttg agacccttat cggcttgaac4620cgctggaata atgccacgtg gaagataatt ccatgaatct tatcgttatc tatgagtgaa4680attgtgtgat ggtggagtgg tgcttgctca ttttacttgc ctggtggact tggccctttc4740cttatgggga atttatattt tacttactat agagctttca tacctttttt ttaccttgga4800
tttagttaat atataatggt atgattcatg aataaaaatg ggaaattttt gaatttgtac4860tgctaaatgc ataagattag gtgaaactgt ggaatatata tttttttcat ttaaaagcaa4920aatttgcctt ttactagaat tataaatata gaaaaatata taacattcaa ataaaaatga4980aaataagaac tttcaaaaaa cagaactatg tttaatgtgt aaagattagt cgcacatcaa5040gtcatctgtt acaatatgtt acaacaagtc ataagcccaa caaagttagc acgtctaaat5100aaactaaaga gtccacgaaa atattacaaa tcataagccc aacaaagtta ttgatcaaaa5160aaaaaaaacg cccaacaaag ctaaacaaag tccaaaaaaa acttctcaag tctccatctt5220cctttatgaa cattgaaaac tatacacaaa acaagtcaga taaatctctt tctgggcctg5280tcttcccaac ctcctacatc acttccctat cggattgaat gttttacttg taccttttcc5340gttgcaatga tattgatagt atgtttgtga aaactaatag ggttaacaat cgaagtcatg5400gaatatggat ttggtccaag attttccgag agctttctag tagaaagccc atcaccagaa5460atttactagt aaaataaatc accaattagg tttcttatta tgtgccaaat tcaatataat5520tatagaggat atttcaaatg aaaacgtatg aatgttatta gtaaatggtc aggtaagaca5580ttaaaaaaat cctacgtcag atattcaact ttaaaaattc gatcagtgtg gaattgtaca5640aaaatttggg atctactata tatatataat gctttacaac acttggattt ttttttggag5700gctggaattt ttaatctaca tatttgtttt ggccatgcac caactcattg tttagtgtaa5760tactttgatt ttgtcaaata tatgtgttcg tgtatatttg tataagaatt tctttgacca5820tatacacaca cacatatata tatatatata tatattatat atcatgcact tttaattgaa5880aaaataatat atatatatat agtgcatttt ttctaacaac catatatgtt gcgattgatc5940tgcaaaaata ctgctagagt aatgaaaaat ataatctatt gctgaaatta tctcagatgt6000taagattttc ttaaagtaaa ttctttcaaa ttttagctaa aagtcttgta ataactaaag6060aataatacac aatctcgacc acggaaaaaa aacacataat aaatttgaat ttcgaccgcg6120gtacccggaa ttgggttata attacctcag gtcgaggaat taattcggta cgtacctaat6180aacttcgtat agcatacatt atacgaagtt atatggatct cgaggcatta cggcattacg6240gcactcgcga gggtcccaat tcgagcatgg agccatttac aattgaatat atcctgccgc6300cgctgccgct ttgcacccgg tggagcttgc atgttggttt ctacgcagaa ctgagccggt6360taggcagata atttccattg agaactgagc catgtgcacc ttccccccaa cacggtgagc6420gacggggcaa cggagtgatc cacatgggac ttttaaacat catccgtcgg atggcgttgc6480gagagaagca gtcgatccgt gagatcagcc gacgcaccgg gcaggcgcgc aacacgatcg6540caaagtattt gaacgcaggt acaatcgagc cgacgttcac ggtaccggaa cgaccaagca6600agctagctta gtaaagccct cgctagattt taatgcggat gttgcgatta cttcgccaac6660tattgcgata acaagaaaaa gccagccttt catgatatat ctcccaattt gtgtagggct6720tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa6780ttatgtgctt agtgcatcta acgcttgagt taagccgcgc cgcgaagcgg cgtcggcttg6840aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga6900caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag6960cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt7020cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca7080acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta7140aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg7200ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat7260caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc7320caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa7380tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa7440ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca7500aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg7560ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg7620atacttcggc gatcaccgct tccctcatga tgtttaactt tgttttaggg cgactgccct7680gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt7740gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga7800aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg7860agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc7920
gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg 7980aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg 8040cattggcggc cttgctgttc ttctacggca agtgctgtgc acggatctgc cctggcttca 8100ggagatcgga agacctcggc cgtccgggcg cttgccggtg gtgctgaccc cggatgaagt 8160ctctagagct ctagagggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg 8220cccagcttct gtatggaacg ggcatgcgga tcagtgaggg tttgcaactg cgggtcaagg 8280atctggattt cgatcacggc acgatcatcg tgcgggaggg caagggctcc aaggatcggg 8340ccttgatgtt acccgagagc ttggcaccca gcctgcgcga gcagggatcg atccaacccc 8400tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcttct gaaaacgaca 8460tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc 8520ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg 8580ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc 8640aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg 8700agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac 8760gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc 8820tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc 8880cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg 8940ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg 9000cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc 9060gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc 9120atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca 9180ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg 9240agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt 9300tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc 9360cgcgcacgtc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct 9420ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg 9480atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt 9540aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc 9600gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc 9660gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg 9720gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag 9780gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct 9840gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac 9900atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga 9960aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag10020gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc10080gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc10140gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt10200aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg10260cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg10320tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag10380gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca10440aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa10500caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc10560gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa10620tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga10680cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc10740acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc10800gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt10860tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc10920cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc10980agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga11040
cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa11100gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg11160gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac11220cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc11280cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga11340gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc11400ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct11460ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat11520ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct11580gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc11640gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg11700tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt11760tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta11820cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa11880agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac11940ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc12000tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc12060tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc12120gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg12180atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag12240cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg12300gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc12360atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt12420aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc12480ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac12540agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa12600ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca12660caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc12720gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata12780cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta12840tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca12900gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga12960cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg13020tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg13080tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg13140caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag13200aaaaaaagga tctcaagaag atccggaaaa cgcaagcgca aagagaaagc aggtagcttg13260cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga13320attgcc 1332權(quán)利要求
1.獲得具有增加的脅迫抗性植物的方法，其包括a.向植物細(xì)胞中引入嵌合基因獲得轉(zhuǎn)基因細(xì)胞，所述嵌合基因含有以下有效連接的DNA片段i.可在植物中表達(dá)的啟動(dòng)子；ii.編碼煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的植物功能酶的DNA區(qū)，所述酶選自煙酰胺酶、煙酸酯磷酸核糖基轉(zhuǎn)移酶、煙酸單核苷酸腺苷轉(zhuǎn)移酶或煙酰胺腺嘌呤二核苷酸合成酶；iii.參與轉(zhuǎn)錄終止和多聚腺苷酸化作用的3’末端區(qū)；b.再生所述的轉(zhuǎn)基因細(xì)胞以獲得轉(zhuǎn)基因植物群；和c.從所述轉(zhuǎn)基因植物群中選擇表現(xiàn)為增加的脅迫抗性的植物或選擇在脅迫條件下與類似的非轉(zhuǎn)基因植物相比表現(xiàn)為降低的活性氧水平或維持高水平NADH的植物。
2.根據(jù)權(quán)利要求1的方法，其中所述DNA區(qū)編碼的酶含有選自SE Q ID No.2、SEQ ID No.4、SEQ ID No.6、SEQ ID No.8、SEQ ID No.10、SEQ ID No.12、SEQ ID No.14、SEQ ID No.16、SEQ ID No.18、SEQ ID No.20、SEQ ID No.22、SEQ ID No.24的氨基酸序列或其編碼的蛋白質(zhì)具有約60％序列同一性并且具有煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑的酶活性。
3.根據(jù)權(quán)利要求1的方法，其中所述DNA區(qū)編碼煙酰胺酶。
4.根據(jù)權(quán)利要求3的方法，其中所述DNA區(qū)編碼的蛋白質(zhì)含有選自SEQ ID No.2、SEQ ID No.12、SEQ ID No.14或SEQID No.16的氨基酸序列。
5.根據(jù)權(quán)利要求3的方法，其中所述DNA區(qū)含有SEQ IDNo.1、SEQ ID No.3、SEQ ID No.11、SEQ ID No.13或SEQ IDNo.15的核苷酸序列。
6.根據(jù)權(quán)利要求1的方法，其中所述DNA區(qū)編碼煙酸酯磷酸核糖基轉(zhuǎn)移酶。
7.根據(jù)權(quán)利要求6的方法，其中所述DNA區(qū)編碼的蛋白質(zhì)含有選自SEQ ID No.4、SEQ ID No.18或SEQ ID No.20的氨基酸序列。
8.根據(jù)權(quán)利要求6的方法，其中所述DNA區(qū)含有SEQ IDNo.3、SEQ ID No.17或SEQ ID No.19的核苷酸序列。
9.根據(jù)權(quán)利要求1的方法，其中所述DNA區(qū)編碼煙酸單核苷酸腺苷轉(zhuǎn)移酶。
10.根據(jù)權(quán)利要求9的方法，其中所述DNA區(qū)編碼的蛋白質(zhì)含有選自SEQ ID No.6、SEQ ID No.8或SEQ ID No.22的氨基酸序列。
11.根據(jù)權(quán)利要求9的方法，其中所述DNA區(qū)含有SEQ IDNo.5、SEQ ID No.7或SEQ ID No.21的核苷酸序列。
12.根據(jù)權(quán)利要求1的方法，其中所述DNA區(qū)編碼NAD合成酶。
13.根據(jù)權(quán)利要求12的方法，其中所述DNA區(qū)編碼的蛋白質(zhì)含有選自SEQ ID No.10或SEQ ID No.24的氨基酸序列。
14.根據(jù)權(quán)利要求12的方法，其中所述DNA區(qū)含有SEQID No.9或SEQ ID No.23的核苷酸序列。
15.根據(jù)權(quán)利要求1-14中任一項(xiàng)的方法，所述方法還包括將所述植物與其他植物雜交的步驟。
16.權(quán)利要求1-14任一項(xiàng)中描述的嵌合基因。
17.含有權(quán)利要求16所述嵌合基因的植物細(xì)胞。
18.含有權(quán)利要求16所述嵌合基因的植物。
19.權(quán)利要求18的植物，其選自棉花、蕓苔屬植物、油料種子油菜、小麥、谷物或玉米、大麥、向日葵、稻、燕麥、甘蔗、大豆、蔬菜、菊苣、萵苣、番茄、煙草、馬鈴薯、甜菜、木瓜、菠蘿、芒果或擬南芥。
20.根據(jù)權(quán)利要求18或19所述的植物，其特征還在于在脅迫條件下與不含這類嵌合基因的類似植物相比，其活性氧水平更低。
21.根據(jù)權(quán)利要求18或19的植物的種子，所述種子含有根據(jù)權(quán)利要求16的嵌合基因。
22.根據(jù)權(quán)利要求16的嵌合基因用于增加植物脅迫抗性的用途。
23.根據(jù)權(quán)利要求16的嵌合基因的用途，所述用途為在脅迫條件下降低植物或植物細(xì)胞中活性氧水平或在脅迫條件下維持植物或植物細(xì)胞中NAD水平。
24.DNA序列增加植物脅迫抗性的用途，所述DNA序列編碼煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的植物功能酶，所述酶選自煙酰胺酶、煙酸酯磷酸核糖基轉(zhuǎn)移酶、煙酸單核苷酸腺苷轉(zhuǎn)移酶或煙酰胺腺嘌呤二核苷酸合成酶。
25.DNA序列降低植物或植物細(xì)胞在脅迫條件下的活性氧水平或維持植物或植物細(xì)胞在脅迫條件下的NAD水平的用途，所述DNA序列編碼煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的植物功能酶，所述酶選自煙酰胺酶、煙酸酯磷酸核糖基轉(zhuǎn)移酶、煙酸單核苷酸腺苷轉(zhuǎn)移酶或煙酰胺腺嘌呤二核苷酸合成酶。
26.DNA序列增加植物脅迫抗性的用途，所述DNA序列編碼的蛋白質(zhì)含有選自SEQ ID No.2、SEQ ID No.4、SEQ IDNo.6、SEQ ID No.8、SEQ ID No.10、SEQ ID No.12、SEQ IDNo.14、SEQ ID No.16、SEQ ID No.18、SEQ ID No.20、SEQ IDNo.22、SEQ ID No.24的氨基酸序列或其編碼的蛋白質(zhì)具有約為60％序列同一性并且具有煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的酶活性。
27.DNA序列降低植物或植物細(xì)胞在脅迫條件下活性氧水平或維持植物或植物細(xì)胞在脅迫條件下NAD水平的用途，所述DNA序列編碼的蛋白質(zhì)含有選自SEQ ID No.2、SEQ ID No.4、SE Q ID No.6、SEQ ID No.8、SEQ ID No.10、SE Q ID No.12、SEQ ID No.14、SEQ ID No.16、SE Q ID No.18、SEQ ID No.20、SEQ ID No.22、SEQ ID No.24的氨基酸序列或其編碼的蛋白質(zhì)具有約為60％序列同一性并且具有煙酰胺腺嘌呤二核苷酸補(bǔ)救合成途徑中的酶活性。
28.DNA序列增加植物脅迫抗性的用途，所述DNA序列含有選自SEQ ID No.1、SEQ ID No.3、SEQ ID No.5、SEQ IDNo.7、SEQ ID No.9、SEQ ID No.11、SEQ ID No.13、SEQ IDNo.15、SEQ ID No.17、SEQ ID No.19、SEQ ID No.21或SEQID No.23的核苷酸序列。
29.DNA序列降低植物或植物細(xì)胞在脅迫條件下的活性氧水平或維持植物或植物細(xì)胞在脅迫條件下的NAD水平的用途，所述DNA序列含有選自SEQ ID No.1、SEQ ID No.3、SEQ IDNo.5、SEQ ID No.7、SEQ ID No.9、SEQ ID No.11、SEQ IDNo.13、SEQ ID No.15、SEQ ID No.17、SEQ ID No.19、SEQ IDNo.21或SEQ ID No.23的核苷酸序列。
全文摘要
通過使用編碼參與NAD補(bǔ)救合成途徑和/或NAD從頭合成途徑的酶的核苷酸序列(如在植物中過表達(dá))使植物和植物細(xì)胞獲得脅迫耐受性。
文檔編號(hào)A01H5/00GK101040049SQ200580032357
公開日2007年9月19日 申請(qǐng)日期2005年9月16日 優(yōu)先權(quán)日2004年9月24日
發(fā)明者M·德布洛克, M·梅茨拉夫, V·格瑟勒 申請(qǐng)人:拜爾生物科學(xué)公司