CN112011565B - 一种棉花转化事件kjc003及其应用 - Google Patents

一种棉花转化事件kjc003及其应用 Download PDF

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CN112011565B
CN112011565B CN202010429529.0A CN202010429529A CN112011565B CN 112011565 B CN112011565 B CN 112011565B CN 202010429529 A CN202010429529 A CN 202010429529A CN 112011565 B CN112011565 B CN 112011565B
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张蕾
罗祖勇
裴睿丽
祁幼林
孟钊红
杨敏
田爱菊
王琪
李莹莹
李秀影
葛玉君
黎跃进
李相敢
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Abstract

本申请公开了一种棉花转化事件KJC003,其特征在于,特征基因序列包含转基因表达盒,所述转基因表达盒的核苷酸序列为SEQ ID NO:9。本发明还提供了用于检测棉花转化事件KJC003的引物对,以及检测方法。本发明的转化事件KJC003的棉花能够同时耐受草甘膦和抵抗鳞翅目害虫的侵染,且结铃、吐絮均不受影响,有利于控制田间杂草,田间害虫,同时提高耕作效率和产量。

Description

一种棉花转化事件KJC003及其应用
技术领域
本申请本发明涉及植物生物技术领域,主要涉及一种用于棉花抗草甘膦和抗鳞翅目害虫的转化事件KJC003及其在棉花育种中的应用。
背景技术
棉花(陆地棉)在世界上许多地区是重要的经济作物,为了生产具有所需性状的棉花,已将生物技术方法应用于这种作物。一种此类所需性状是除草剂耐受性,一种此类所需性状是抗虫性。除草剂耐受性转基因在植物中的表达能够赋予植物所需的除草剂耐受性状,抗虫性转基因在植物中的表达课赋予植物所需的抗虫性状。许多不同的因素影响转基因的表达,但是转基因的表达可能受到转基因插入的染色体位置和基因组结果的影响。例如,已经在植物中观察到,在转基因的染色体插入位点方面不同但在其他方面都相同的单个事件之间,通常存在着转基因表达水平和模式的变异。在事件之间还可能存在不想要的和/或想要的表型或农学差异。因此,为了选择具有所需性状和使其适合于商业化目的所需的最适表型和农业特性两者的事件,通常需要产生并分析大量单个植物转化事件。这样的选择通常需要经过多年、在多个地点并在多种不同条件下对许多事件进行温室和田间试验,以便可以收集到大量农艺学、表型和分子数据。然后必须由科学家和农学家对得到的数据进行分析,其目的是选择商业上适合的事件。这样的事件一旦被选择,随后可能用于使用植物育种方法将所需性状渗入到其他遗传背景中,并由此产生含有所需性状并适当地适应于特定当地生长条件的大量不同作物品种,这些作物品种含有所需性状并且适当的适应特定的当地农艺条件。
发明内容
在研究过程中,申请人发现了一个可使棉花获得抗草甘膦性状和抗虫(鳞翅目)的转基因表达盒,转基因的棉花植株在高剂量草甘膦和鳞翅目害虫的侵染的情况下仍正常生长。
本发明的一个方面在于提供一种棉花抗草甘膦和抗虫转化事件 KJC003,其特征基因序列包含转基因表达盒,所述转基因表达盒的核苷酸序列为SEQ ID NO:9。
在根据本发明的一个实施方案中,所述特征基因序列还包含5'端连接序列,所述5'连接序列选自SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5或 SEQ ID NO:7中的任一个。
在根据本发明的一个实施方案中,所述特征基因序列还包含3'端连接序列,所述3'端连接序列选自SEQ ID NO:2、SEQ ID NO:4、SEQ ID NO:6和 SEQ ID NO:8中的任一个。
在根据本发明的一个实施方案中,所述特征基因序列为核苷酸序列为SEQ ID NO:10的基因序列或其互补序列。
在根据本发明的一个实施方案中,所述特征序列位于四倍体棉花的 D基因组第8组染色体上。
本发明的另一方面在于提供用于检测上述的棉花转化事件KJC003的引物对,所述引物对包含特异性识别侧翼序列的第一引物对和/或第二引物对,所述第一引物对由SEQID NO:11和SEQ ID NO:12(P681:5’- CCAGAATCCTTGTCAGATT-3’和D08-3242F:5’-TACGGCTTATCACCCTCA-3’) 组成;所述第二引物对由SEQ ID NO:13和SEQ ID NO:14组成。FMV1:5’ -ATAAAGGCAAAGATGCTTTTGGTAGGTG-3’和D08-277R:5’- TTTCCTTGTGGTCAGTTT-3’。
本发明进一步提供了一种鉴定棉花生物样品中棉花转化事件KJC003 的方法,其包括:(a)从待鉴定的棉花生物样品提取DNA样品;(b) 以提取的DNA样品为模板,使用第一引物对(SEQ ID NO:11和SEQ ID NO:12) 和第二引物对(SEQ ID NO:13和SEQ ID NO:14)进行PCR扩增;(c)检测 PCR扩增产物的长度与棉花转化事件KJC003中的理论核苷酸序列长度比较。若以第一引物对扩增得到的扩增产物的核苷酸序列长度为1177bp,或以第二引物扩增等到的扩增产物的核苷酸序列长度为614bp,则表明样品中存在棉花抗草甘膦抗虫转化事件KJC003。
进一步的,上述用途包括具备棉花抗草甘膦抗虫转化事件KJC003的棉花植株、棉花植物细胞、种子、子代植株、杂交植株、棉油、棉花蛋白质、棉花商品产品等等。
本发明的另一方面在于提供上述的转化事件KJC003以提高棉花对草甘膦和鳞翅目害虫的抗性,进行棉花育种或用作分子标记的用途。
附图说明
图1所示为棉花抗草甘膦抗虫转化事件KJC003中基因表达盒在包含事件KJC003的棉花植物的基因组中的结构。
图2为棉花抗草甘膦抗虫转化事件KJC003 T2代抗草甘膦的测试,其中,右侧为对照组的非转基因新陆早45,左侧为本发明的抗草甘膦抗虫转化事件KJC003 T2代棉花。对照株和实验株在棉花4叶期分别喷施浓度 1×(840g ae/ha)草甘膦,喷施10天后观察,拍照,全生育期跟踪棉花生长、开花、结铃、吐絮情况。
图3为棉花抗草甘膦抗虫转化事件KJC003 T2代抗棉铃虫测试,其中,测试用虫为初孵的棉铃虫。左侧为对照组的非转基因新陆早45,右侧为本发明的抗草甘膦抗虫转化事件KJC003 T2代棉花。
图4为棉花抗草甘膦抗虫转化事件KJC003 T2代抗小地老虎测试,其中,测试用虫为初孵的小地老虎。左侧为对照组的非转基因新陆早45,右侧为本发明的抗草甘膦抗虫转化事件KJC003 T2代棉花。
图5为棉花抗草甘膦抗虫转化事件KJC003 T2代的接虫试验。左侧为对照组的非转基因新陆早45,右侧为本发明的抗草甘膦抗虫转化事件 KJC003 T2代棉花。
图6为棉花抗草甘膦转化事件KJC003的检测结果图,其中,M:Trans 2K plus II,分子量标准从上到下依次为8kb、5kb、3kb、2kb、1kb、750bp、 500bp、250bp、100bp;泳道1和2以KJC003 T2代的2个单株DNA为模板扩增产物(1177bp),泳道3以质粒DNA为模板扩增产物,泳道4 以新陆早45DNA为对照模板扩增产物,泳道5以ddH2O为模板PCR产物;
图7为棉花抗草甘膦转化事件KJC003的检测结果图,其中,M:Trans 2K plus II,分子量标准从上到下依次为8kb、5kb、3kb、2kb、1kb、750bp、 500bp、250bp、100bp;泳道1和2以KJC003 T2代的2个单株DNA为模板扩增产物(614bp),泳道3以质粒DNA为模板扩增产物,泳道4以新陆早45DNA为对照模板扩增产物,泳道5以ddH2O为模板PCR产物;
具体实施方式
以下实施例用于说明本申请,但不用来限制本申请的范围。
下面将更详细地描述本申请的具体实施例。提供这些实施例是为了能够更透彻地理解本申请,并且能够将本申请的范围完整的传达给本领域的技术人员。
如在通篇说明书及权利要求当中所提及的“包含”或“包括”为一开放式用语,故应解释成“包含但不限定于”。说明书后续描述为实施本申请的较佳实施方式,然所述描述乃以说明书的一般原则为目的,并非用以限定本申请的范围。本申请的保护范围当视所附权利要求所界定者为准。
“引物”通常是高度纯化的分离的多核苷酸,其被设计用于涉及热扩增的特异性退火或杂交方法中。一对引物可以与模板DNA例如棉花基因组DNA样品一起使用在热扩增例如聚合酶链反应(PCR)中,以产生扩增子,其中从这样的反应产生的扩增子具有与位于引物杂交到模板处的两个位点之间的模板DNA的序列相对应的DNA序列。当在本文中使用时,“扩增子”是使用扩增技术合成的DNA碎片或片段。本发明的扩增子包含至少一个以SEQ ID NO:1-10提供的序列。引物通常被设计成与互补的靶DNA链杂交以在引物与靶DNA链之间形成杂交体,并且引物的存在是聚合酶使用靶DNA链作为模板开始引物延伸(即另外的核苷酸聚合到伸长的核苷酸分子中)的识别位点。当在本发明中使用时,引物对意图指使用结合双链核苷酸片段的相反链的两个引物以通常在热扩增反应或其他常规核酸扩增方法中实现线性扩增由引物对的单个成员的结合所靶定的位置之间的多核苷酸片段的目的。
本文中术语“分离的”是指分子至少部分地与在天源或自然状态下通常与其相伴的其他分子分离。在一种实施方式中,术语“分离的”是指DNA分子与在天源或自然状态下通常邻接所述DNA分子的核酸至少部分地分离。因此,例如作为重组技术的结果,与正常情况下不与其相关的调节或编码序列融合的DNA分子在本文中被认为是分离的。即使在整合到宿主细胞的染色体中或与其他DNA分子一起存在于核酸溶液中时,这样的分子也被认为是分离的。
可以使用本领域技术人员公知的大量方法来分离和操作本发明中公开的DNA分子或其片段。例如,可以使用PCR(聚合酶链反应)技术来扩增特定的起始DNA分子和/或产生原始分子的变体。DNA分子或其片段也可以通过其他技术来获得,例如通过使用化学手段直接合成所述片段,正如通常通过使用自动化寡核苷酸合成仪所实现的。
下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的材料、试剂、仪器等,如无特殊说明,均为自常规生化试剂商店购买得到的。
实施例1:棉花的转化和KJC003事件选择
本实施例描述了转化事件KJC003的产生、分析和选择。
耐受草甘膦和抗鳞翅目害虫侵染的转基因棉花转化事件KJC003利用包含图1中示出的表达盒的植物转化载体,通过土壤杆菌介导的棉花转化产生。为了产生KJC003事件,将新陆早45棉花材料用于植物转化。将棉花细胞转化并再生成完整棉花植物。选择具有正常表型特征的生根植物,并将其转移至土壤进行生长和进一步评估。
将T0植物转移至土壤,PCR鉴定并选择含T-DNA表达盒的T0植物。T-DNA I表达盒含有P-FMV tsf1启动子,该启动子包含:玄参花叶病毒(FMV)增强子,拟南芥transcriptionfactor 1启动子及其5’UTR调控序列;可操作地与编码来自于拟南芥的5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)的叶绿体转运肽的N-端叶绿体转运肽的DNA分子(CTP2) 连接;可操作地与编码来自于土壤农杆菌CP4的5-烯醇式丙酮酰莽草酸 -3-磷酸合酶(EPSPS)连接;可操作地与豌豆的rbcs2 E9基因3’UTR DNA 分子(T-E9)连接;P-FMV tsf1启动子与拟南芥启动子Action 2连接,拟南芥启动子Action 2包含Action2基因启动子及其5’UTR调控序列;可操作地与编码Vip3Aa蛋白的基因连接;可操作地与编码胭脂碱合成酶基因的终止子连接(T-nos);可操作地与含有增强子的烟草花叶病毒启动子 CaMV35S连接,与编码抗虫蛋白Cry1Ac连接,可操作地与7S终止子连接。 T-DNA I的RB与T-DNA II的LB相邻。T-DNA II含有P-FMV tsf1启动子,该启动子包含:玄参花叶病毒(FMV)增强子,拟南芥transcriptionfactor 1启动子及其5’UTR调控序列;可操作地与编码来自于拟南芥的5-烯醇式丙酮酰莽草酸-3-磷酸合酶(EPSPS)的叶绿体转运肽的N-端叶绿体转运肽的DNA分子(CTP2)连接;可操作地与编码氨基糖苷腺苷酰转移酶基因 (aadA)连接;可操作地与豌豆的rbcs2 E9基因3’UTR DNA分子(T-E9) 连接。
以草甘膦1×(840g ae/ha)的田间施用量进行除草剂筛选T1代棉花,抗性单株自交收取种子。通过温室接棉铃虫试验和室内生测,筛选到最抗的转化事件KJC003。T2代Taqman分析CP4-EPSPS、Vip3Aa、Cry1Ac和aadA 均为单拷贝插入。
实施例2:转化事件KJC003序列的表征
使用包括植物表达盒的完整编码区及其相应的调控元件、启动子、内含子和多腺苷化序列的分子DNA探针。分析显示,KJC003事件含有转基因DNA均为单拷贝插入。进行Hi-Tail PCR和DNA序列分析以确定插入片段-植物基因组的5’和3’接合部,证实插入片段内元件的组织(图1),并确定包含转化事件KJC003的棉花植物中插入片段的完整 DNA序列(在本文中提供为SEQ ID NO:9)。在基因组中包含图1中示出的连接的转基因遗传元件并且对草甘膦除草剂抗性和对鳞翅目害虫有抗性的棉花植物是本发明的一个方面。
SEQ ID NO:1是30个核苷酸的序列,其表示棉花基因组DNA与整合的转基因表达盒的5’连接区域。SEQ ID NO:1位于SEQ ID NO:10中747—776 核苷酸位置处。
SEQ ID NO:2是30个核苷酸的序列,其表示棉花基因组DNA与整合的转基因表达盒的3’连接区域。SEQ ID NO:2位于SEQ ID NO:10中 17943—17972核苷酸位置处。
SEQ ID NO:3是60个核苷酸的序列,其表示棉花基因组DNA与整合的转基因表达盒的5’连接区域。
SEQ ID NO:4是60个核苷酸的序列,其表示棉花基因组DNA与整合的转基因表达盒的3’连接区域。
SEQ ID NO:5是100个核苷酸的序列,其表示棉花基因组DNA与整合的转基因表达盒的5’连接区域。
SEQ ID NO:6是100个核苷酸的序列,其表示棉花基因组DNA与整合的转基因表达盒的3’连接区域。
SEQ ID NO:7是侧邻KJC003的插入DNA直到并包括一部分整合的转基因表达盒的5’序列。
SEQ ID NO:8是侧邻KJC003的插入DNA直到并包括一部分整合的转基因表达盒的3’序列。
SEQ ID NO:9是整合的转基因表达盒的序列。
SEQ ID NO:10是侧邻插入DNA的5’序列(SEQ ID NO:7)、整合的转基因表达盒(SEQID NO:9)和侧邻插入DNA的3’序列(SEQ ID NO:8)的连续核苷酸序列。SEQ ID NO:10包含SEQ ID NO:1-9。
实施例3.棉花转化事件KJC003筛选鉴定
1.棉花抗草甘膦抗虫转化事件KJC003 T1代抗草甘膦测试:
试验时间:2018年12月,试验地点:海南三亚崖州区南滨,测试对照为非转基因受体新陆早45,于棉花4叶期喷施浓度1×草甘膦(41%草甘膦异丙胺盐水剂,840g ae/ha),喷施10天后记录、拍照,在全生育期跟踪棉花,生长,开花,结铃和吐絮情况。
试验结果表明,4叶期喷洒1×草甘膦(840g ae/ha),棉花抗草甘膦抗虫转化事件KJC003 T1代植株叶片无坏死斑,生长点正常,叶色浓绿,表明植株对1倍浓度草甘膦有较好的抗性,如图2所示。
2.棉花抗草甘膦和抗虫的转化事件KJC003 T2代抗虫的测试:
转基因棉花抗虫试验的测试参照国际标准GB/T 22101.1-2008进行。
a)抗虫生测试验-棉铃虫
试验时间:2019年7月,试验地点:公司实验室光照培养箱中,测试对照为新陆早45,测试用虫为初孵的棉铃虫。
将棉叶取样于培养皿大小,每个皿中接初孵幼虫5头,重复3次,皿底放置1张湿滤纸,保持棉叶在供试期内不干。接虫后用封口膜将双重皿封严,防止幼虫逃逸和保持皿内湿度。放入25-28℃的养虫室中饲养,每天光照饲养14小时,黑暗饲养10小时(L:D=14:10)。
在接虫后的第三天和第六天分别统计幼虫死亡数和活虫数,计算幼虫死亡率。测试的结果如图3所示。室内生测棉铃虫的校正死亡率为100%,转基因事件KJC003的抗虫级别为高抗。
b)抗虫生测试验-小地老虎
试验时间:2019年7月,试验地点:公司实验室光照培养箱中,测试对照为新陆早45,测试用虫为初孵的小地老虎。
取培养皿大小的棉叶,每个皿中接初孵幼虫5头,重复3次,皿底放置1张湿滤纸,保持棉叶在供试期内不干。接虫后用封口膜将双重皿封严,防止幼虫逃逸和保持皿内湿度。放入25-28℃的养虫室中饲养,每天光照饲养14小时,黑暗饲养10小时(L:D=14:10)。
在接虫后的第三天和第六天分别统计幼虫死亡数和活虫数,计算幼虫死亡率。测试的结果如图4所示。室内生测小地老虎的校正死亡率为100%,转基因事件KJC003的抗虫级别为高抗。
c)大田的接虫试验
试验时间:2019年7月,试验地点:北京市昌平区百善镇试验大棚内,测试对照为新陆早45,供试虫源为人工饲料饲喂2天的1龄棉铃虫。在棉花的盛花期,使用孵出后饲喂2天的棉铃虫幼虫,每个事件接虫 10株,每株5头。接虫15天后,统计被害蕾铃。测试的结果如图5所示。接虫试验蕾铃受寒减退率为100%,转基因事件KJC003对棉铃虫的抗虫级别为高抗。
实施例4.Taqman分析
用本领域常用的植物DNA提取方法(李可凡.一种棉花DNA快速高效简便的提取方法[J].江苏农业科学,2015,000(005):54-55,56)提取含有转化事件KJC003的棉花材料的基因组DNA,在50ul水中溶解。取1ul为模板DNA,分别以(SEQ ID NO:15)5’-GGCAGCCTTCGTATCGGAG-3’和 (SEQ ID NO:16)5’-CTCGTGTCGGAAAACCCTGT-3’为引物,(SEQID NO:17) FAM-CAG GTCCATGAACTCCGGGAAGCTC-BHQ1为探针,检测CP4-EPSPS拷贝数;以(SEQ ID NO:18)5’-CTGAAGTGTCCCAAGAGGTTAGAGTC-3’和(SEQ ID NO:19)5’-GTTGTTCTCGATCTCGTGGATG-3’为引物,(SEQ ID NO:20) FAM-CAGCCGCTTACAAGGAGGGATACGGTGAGG-BHQ1为探针,检测Cry1Ac 拷贝数;以(SEQ ID NO:21)5’-TCCACTACGAGGACACCAACAAC-3’和(SEQ ID NO:22)5’-CCGTTCTGGCTCTTCAGGATC-3’为引物,(SEQ ID NO:23) FAM-TGGAGGACTACCAGACCATCAACAAGCG-BHQ1为探针,检测Vip3Aa拷贝数;以(SEQ ID NO:24)5’-AGAATGGCAGCGCAATGAC-3’和(SEQ ID NO:25) 5’-CAGCAAGATAGCCAGATCAATGTC-3’为引物,(SEQ ID NO:26) FAM-CTTGCAGGTATCTTCGAGCCAGCCAC-BHQ1为探针,检测aadA拷贝数。经过分析,转化事件KJC003为单拷贝。
实施例5.侧翼序列分析
按照参考文献(Liu Y G,et al.,2007)的Hi-TailPCR方法,获得KJC003 事件插入片段的侧翼序列。在需要Tail PCR位置的大约100bp,200bp, 300bp处各设计一条特异性引物,FMV-3(SEQ ID NO:27):5’ -AACTTACTGACCATGATGCATGTGCTGG-3’,FMV-2(SEQ ID NO:28):5’ -ACGATGGACTCCAGTCCCAGCTGCTCGATGTTGACAAGATTAC-3’,FMV-1(SEQ ID NO:13):5’-ATAAAGGCAAAGATGCTTTTGGTAGGTG-3’,简并引物同文献(Liu Y G,Chen Y.High-efficiency thermal asymmetric interlaced PCR for amplification of unknownflanking sequences[J].BioTechniques,2007, 43(5):649-656)。
Hi-Tail PCR扩增体系以及扩增流程
表1扩增体系:
Figure BDA0002499998450000101
Figure BDA0002499998450000111
表2扩增程序:
Figure BDA0002499998450000112
Figure BDA0002499998450000121
Tail II PCR和Tail III PCR的产物各吸取2ul加2ul 6×Loading Buffer跑1%琼脂糖凝胶电泳。挑选长度大于已知且Tail II PCR比Tail III PCR产物大的目的条带进行测序。利用公布的棉花(Gossypium hirsutum)基因组序列 (https://www.cottongen.org/)Gossypium hirsutum(AD1)TM-1genome ZJU-Improved v2.1_a1与测序结果进行分析比对,发现与D08染色体上的序列高度同源。
然后根据基因组序列在该位点的上下游800bp范围内设计引物 D08-3242F,D08-277R。运用p681(SEQ ID NO:11)和D08-3242F(SEQ ID NO:12),FMV1(SEQ ID NO:13)和D08-277R(SEQ ID NO:14)分别扩增样品,然后对产物进行测序,获得与插入序列接合的棉花侧翼DNA序列。
实施例6.染色体定位
根据所获得的棉花侧翼序列,利用公布的棉花(Gossypium hirsutum) 基因组序列(https://www.cottongen.org/)Gossypium hirsutum(AD1)TM-1 genome ZJU-Improvedv2.1_a1进行对比分析,可知转化事件KJC003中,与插入序列接合的棉花侧翼DNA序列与D08染色体上的序列高度同源,因此可知,转化事件KJC003中外源DNA插入序列的整合位点位于受体四倍体棉花的D基因组第8组染色体上。
实施例7.转化事件KJC003检测
使用DNA引物对进行PCR扩增以检测KJC003事件,以转化事件 KJC003的DNA为模板,以p681(SEQ ID NO:11)和D08-3242F(SEQ ID NO:12),为引物,55℃退火,72℃延伸0.5min,华大2ⅹSuper PCR Mix扩增得到 T-E9到LB侧翼序列片段1177bp。以KJC003事件DNA为模板,FMV1(SEQ ID NO:13)和D08-277R(SEQ ID NO:14)为引物,55℃退火,72℃延伸1min,华大2ⅹSuper PCR Mix扩增得到P-FMV tsf1到RB侧翼序列片段614bp。
利用上述引物对进行PCR鉴定使用的是本领域常用的方法。利用引物对分别扩增转化事件KJC003、新陆早45的棉花样品的结果如图6和图7所示。
虽然,上文中已经用一般性说明及具体实施例对本申请作了详尽的描述,但在本申请基础上,可以对至作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本申请精神的基础上所做的这些修改或改进,均属于本申请要求保护的范围。
SEQ ID No.1:AATAAATAAA TAAATCAATG TACTTTTCAT
SEQ ID No.2:AACTATCAGT GTTTGCAATT ATTTACCTAC
SEQ ID No.3:TTTAGGTTGT AAATAAATAA ATAAATAAAT CAATGTACTT TTCATGTCCGGGAAATCTAC
SEQ ID No.4:CCCGCCTTCA GTTTAAACTA TCAGTGTTTG CAATTATTTA CCTACAAATAAAAAAACCAA
SEQ ID No.5:
Figure BDA0002499998450000141
SEQ ID No.6:
Figure BDA0002499998450000142
SEQ ID No.7:
Figure BDA0002499998450000143
SEQ ID No.8:
Figure BDA0002499998450000144
SEQ ID No.9:
Figure BDA0002499998450000145
Figure BDA0002499998450000151
Figure BDA0002499998450000161
Figure BDA0002499998450000171
Figure BDA0002499998450000181
Figure BDA0002499998450000191
Figure BDA0002499998450000201
Figure BDA0002499998450000211
SEQ ID No.10:
Figure BDA0002499998450000212
Figure BDA0002499998450000221
Figure BDA0002499998450000231
Figure BDA0002499998450000241
Figure BDA0002499998450000251
Figure BDA0002499998450000261
Figure BDA0002499998450000271
Figure BDA0002499998450000281
SEQ ID NO:11
ccagaatccttgtcagatt
SEQ ID NO:12
tacggcttatcaccctca
SEQ ID NO:13
ataaaggcaaagatgcttttggtaggtg
SEQ ID NO:14
tttccttgtggtcagttt
SEQ ID NO:15
GGCAGCCTTCGTATCGGAG
SEQ ID NO:16
CTCGTGTCGGAAAACCCTGT
SEQ ID NO:17
GTCCATGAACTCCGGGAAGCTC
SEQ ID NO:18
CTGAAGTGTCCCAAGAGGTTAGAGTC
SEQ ID NO:19
GTTGTTCTCGATCTCGTGGATG
SEQ ID NO:20
CCGCTTACAAGGAGGGATACGGTGAGG
SEQ ID NO:21
TCCACTACGAGGACACCAACAAC
SEQ ID NO:22
CCGTTCTGGCTCTTCAGGATC
SEQ ID NO:23
TGGAGGACTACCAGACCATCAACAAGCG
SEQ ID NO:24
AGAATGGCAGCGCAATGAC
SEQ ID NO:25
CAGCAAGATAGCCAGATCAATGTC
SEQ ID NO:26
CTTGCAGGTATCTTCGAGCCAGCCAC
SEQ ID NO:27
AACTTACTGACCATGATGCATGTGCTGG
SEQ ID NO:28
ACGATGGACTCCAGTCCCAGCTGCTCGATGTTGACAAGATTAC。
序列表
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<120> 一种棉花转化事件KJC003及其应用
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actgtgggcg taccgtacag cttataaagg acccataggt atgtctcctt atcgactaat 180
ttttggtaaa ccgtgtcatc ttcctgttga actagaacat aaagcgtttc gggaggtcaa 240
gcaatgcaac atggagttgg aagttgcagg taagtatcgt cagttatata ttcaggaact 300
tgaagaaatt caacgagaag cgtatgaaag tgcccaaatt tataaggata agatcaaggc 360
ataccatgat cagaagatat cctgtaaata attttcggta cgacataaag tattattcta 420
tgaccctaca ttaagaattt ttgcaggtaa gcttcaaact aagtggttaa ggcattttgt 480
tattactcac gtatttccac atggtacagt cgaggttaaa agcgaaaaat ctgcaaaaaa 540
tttcaaggtc aatggacagc gattaaagtc tttctacgaa aattttcaag tccacatggt 600
cgaggaatta gtcctcgaag agccggttaa ataaatttcc aggtcgtcga gctaatgacg 660
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caattattta cctacaaata aaaaaaccaa catccctaac cttaaaaccc caaaacccaa 60
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gacctataca aaaaaaaccc caatacccaa aacaccaact agcccaatat ggccccaaaa 180
aatgaaaagg aaggaaattg ggccgaacga cctagctggc ccaaagaaaa aaaaagaaca 240
agaaatggac aagaggccca aactgaccac aaggaaaaaa aaagagaaaa gagaaaaaaa 300
gaaaacgaaa aaataaaaca cccaaaatct ttaaaaccct aaaccccttt tttcttctcc 360
atctttttca aaacaaaacc ctaaccccac actacaccgc tgccacctca accatcgtta 420
gagtcgtcgt cgagtgttgt caccaccatc atcgtccaca tttgtcgtcg agcgctcacg 480
agtcaccatg gtctcgtgag ccaccattgt cgaacgtcat ttttcagagt atcgttgctc 540
accccattgc cctatttcat catcgtcgaa cttagacacc accacgttga cgcggcatcg 600
ccgttttaca ccatcaccaa cacgtcgtcg ctccgtcctt cctcaaacac catctcatct 660
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caatgtactt ttcatgtccg ggaaatctac atggatcagc aatgagtatg atggtcaata 60
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gcgttattat aaaatgaaag tacattttga taaaacgaca aattacgatc cgtcgtattt 180
ataggcgaaa gcaataaaca aattattcta attcggaaat ctttatttcg acgtgtctac 240
attcacgtcc aaatgggggc ttagatgagg cgcgccggta ccgtcgacat tgatgcatgt 300
tgtcaatcaa ttggcaagtc ataaaatgca ttaaaaaata ttttcatact caactacaaa 360
tccatgagta taactataat tataaagcaa tgattagaat ctgacaagga ttctggaaaa 420
ttacataaag gaaagttcat aaatgtctaa aacacaagag gacatacttg tattcagtaa 480
catttgcagc ttttctaggt ctgaaaatat atttgttgcc tagtgaataa gcataatggt 540
acaactacaa gtgttttact cctcatatta acttcggtca ttagaggcca cgatttgaca 600
catttttact caaaacaaaa tgtttgcata tctcttataa tttcaaattc aacacacaac 660
aaataagaga aaaaacaaat aatattaatt tgagaatgaa caaaaggacc atatcattca 720
ttaactcttc tccatccatt tccatttcac agttcgatag cgaaaaccga ataaaaaaca 780
cagtaaatta caagcacaac aaatggtaca agaaaaacag ttttcccaat gccataatac 840
tcaaactcag taggattctg gtgtgtgcgc aatgaaactg atgcattgaa cttgacgaac 900
gttgtcgaaa ccgatgatac gaacgaaagc tgaattctca ggcagccttc gtatcggaga 960
gttcgatctt cgcgcccagc ccggccatca ggtccatgaa ctccgggaag ctcgtggcga 1020
tcatcgtggc atcgtccacc gtgacagggt tttccgacac gaggcccatg acgaggaagc 1080
tcatggcgat gcggtgatcg agatgggtgg cgacggcggc gcccgaggcg ttgccgagcc 1140
ccttgccgtc agggcggcca cgcacgacga gcgacgtctc gccctcatcg caatccacgc 1200
cattgagctt gaggccattg gcgacggccg agaggcggtc gctttccttg acgcggagtt 1260
cttccagacc gttcatcacg gtcgcccctt ccgcgaaggc ggcggcgaca gcgagaatcg 1320
gatattcgtc gatcatcgaa ggcgcgcggt cttccggcac cgtgacgccc ttcagcgtgg 1380
aggagcgaac gcgcaggtcc gccacgtctt cgccgccggc aaggcgcggg ttgatgactt 1440
cgatgtcggc gcccatttcc tgcagcgtca ggatgaggcc ggtgcgggtg gggttcatca 1500
gcacgttgag gatggtgacg tcggagcccg gaacaagcag ggccgcaacc agcgggaagg 1560
ccgtcgagga cgggtcgccc ggcacgtcga tgacttggcc ggtgagcttg ccgcggcctt 1620
ccaggcggat ggtgcgcacg ccgtccgcat ccgtctcgac ggtaaggttg gcgccaaagc 1680
cctgcagcat cttttccgta tgatcgcgcg tcatgatcgg ctcgatgacc gtcgtgatgc 1740
cgggcgtgtt gaggccggcg agcagcacgg cggacttcac ctgtgcggag gccatcggca 1800
cgcggtaggt gatcggcgtc ggcgtcttcg gcccgcgcaa ggtaacggga agacggtcac 1860
cgtcttccga tttcacctgc acgcccattt cgcgcagcgg gttcaacacg cggcccatcg 1920
ggcgctttgt gagcgaggcg tcgccgatga aggtgctgtc gaaatcgtag accccgacga 1980
ggcccatcgt caggcggcag cccgtggcgg cattgccgaa atcgagcggc gcctcaggcg 2040
ccaggaggcc gccattgccg acgccatcga tgatccaggt gtcgccttcc ttacggatgc 2100
gggcgcccat cgcctgcatg gccttgcccg tattgatgac gtcctcgcct tccagaaggc 2160
cggtgatgcg cgtttcaccg ctcgcgagac cgccgaacat gaaggaccgg tgggagatcg 2220
acttgtcgcc gggaatgcgg acggttccgg aaaggccaga ggatttgcgg gcggttgcgg 2280
gccggctgct tgcaccgtga agcatgcacg ccgtggaaac agaagacatg accttaagag 2340
gacgaagctc agagccaatt aacgtcatcc cactcttctt caatccccac gacgacgaaa 2400
tcggataagc tcgtggatgc tgctgcgtct tcagagaaac cgataaggga gatttgcgtt 2460
gactggattt cgagagattg gagataagag atgggttctg cacaccattg cagattctgc 2520
taacttgcgc catggtagtt ctctggtcaa caaatctgca aaaaaaagaa caatcaaata 2580
ttgaatcaat aaaaacagta gacatatata accattatga aaatcatctt taacacaaaa 2640
caatcatagt aaaaagccgc tatcggagtc aaaccgattt ctaccacaaa aattgtggtg 2700
ctctatcatc tgagctaagc gagctaaatt caatcaaatc ggcaatttca agagatataa 2760
agcaaacaat ccgagaatta cccttagaaa aagggaacac tggatttgtt atgctactcc 2820
agaattaccc aatcacaatt caaatcgaca tacatcttaa atacgcaaat caacaagttc 2880
actcgtatat agatccacca accaacaatc aatcgaacaa attaaaatct caaaacaaga 2940
tcgatcagaa tgggaaaata gtttcatgaa taaattatcg tttagaagca gtgaaacgtt 3000
gatattaaca atcgataaaa tctaaacaac agatcagaat tttcagaacg atcaatcaac 3060
aaagagcatc gaatccaaca atcatcgaaa taatatgagc aacgatcagc agacgaaaaa 3120
gaaaaagaac gatctaaggg tacgttaacg cttacctcag agatatcgca gaaaagtgtg 3180
tagcggctag ggtcaataag aacgataaat ggaatcagca acgagagtgg ttttatttat 3240
agcgttgaat ccgaaaccaa tgttagggtt taacataaaa taatatcatt ctttaaatta 3300
ccattatgcc cctgtttagc cgtgaatact caatacctct tgggttggtt gtgttttatg 3360
aatgtttagt actacatgtt acaaacatca tttaatcaca ccttttaatt aaggttattc 3420
aagcaaatgg ttatacatgt tcgtctccca aaaatttcat tgattgtgga ccatcaatct 3480
atatgacact cttctctaga gtaaatattt gatatctcaa tgaatgcaga ttatcaagag 3540
ccatttatat catttattca aacaccaaat aaaataatga acaagtaaag tatatattaa 3600
cctcaaaatc tcatgacaat ggaaatggtt tattataaca accatatgtg tcccattcca 3660
cgagcaacct ccctcaagag agaaacttcc agatctcttc cttacaaacc tttgaggatg 3720
ggagttcctt cttggttttg gcgataccaa tttgaataaa gtgatatggc tcgtaccttg 3780
ttgattgaac ccaatctgga atgctgctaa atcctgagct caagctaatt cttttgtggt 3840
cgtcactgcg ttcgtcatac gcattagtga gtgggctgtc aggacagctc ttttccacgt 3900
tattttgttc cccacttgta ctagaggaat ctgctttatc tttgcaataa aggcaaagat 3960
gcttttggta ggtgcgccta acaattctgc accattcctt ttttgtctgg tccccacaag 4020
ccagctgctc gatgttgaca agattacttt caaagatgcc cactaacttt aagtcttcgg 4080
tggatgtctt tttctgaaac ttactgacca tgatgcatgt gctggaacag tagtttactt 4140
tgattgaaga ttcttcattg atctcctgta gcttttggct aatggtttgg agactctgta 4200
ccctgacctt gttgaggctt tggactgaga attaagcttg tcgacaaaat ttagaacgaa 4260
cttaattatg atctcaaata cattgataca tatctcatct agatctaggt tatcattatg 4320
taagaaagtt ttgacgaata tggcacgaca aaatggctag actcgatgta attggtatct 4380
caactcaaca ttatacttat accaaacatt agttagacaa aatttaaaca actatttttt 4440
atgtatgcaa gagtcagcat atgtataatt gattcagaat cgttttgacg agttcggatg 4500
tagtagtagc cattatttaa tgtacatact aatcgtgaat agtgaatatg atgaaacatt 4560
gtatcttatt gtataaatat ccataaacac atcatgaaag acactttctt tcacggtctg 4620
aattaattat gatacaattc taatagaaaa cgaattaaat tacgttgaat tgtatgaaat 4680
ctaattgaac aagccaacca cgacgacgac taacgttgcc tggattgact cggtttaagt 4740
taaccactaa aaaaacggag ctgtcatgta acacgcggat cgagcaggtc acagtcatga 4800
agccatcaaa gcaaaagaac taatccaagg gctgagatga ttaattagtt taaaaattag 4860
ttaacacgag ggaaaaggct gtctgacagc caggtcacgt tatctttacc tgtggtcgaa 4920
atgattcgtg tctgtcgatt ttaattattt ttttgaaagg ccgaaaataa agttgtaaga 4980
gataaacccg cctatataaa ttcatatatt ttcctctccg ctttgaattg tctcgttgtc 5040
ctcctcactt tcatcagccg ttttgaatct ccggcgactt gacagagaag aacaaggaag 5100
aagactaaga gagaaagtaa gagataatcc aggagattca ttctccgttt tgaatcttcc 5160
tcaatctcat cttcttccgc tctttctttc caaggtaata ggaactttct ggatctactt 5220
tatttgctgg atctcgatct tgttttctca atttccttga gatctggaat tcgtttaatt 5280
tggatctgtg aacctccact aaatcttttg gttttactag aatcgatcta agttgaccga 5340
tcagttagct cgattatagc taccagaatt tggcttgacc ttgatggaga gatccatgtt 5400
catgttacct gggaaatgat ttgtatatgt gaattgaaat ctgaactgtt gaagttagat 5460
tgaatctgaa cactgtcaat gttagattga atctgaacac tgtttaaggt tagatgaagt 5520
ttgtgtatag attcttcgaa actttaggat ttgtagtgtc gtacgttgaa cagaaagcta 5580
tttctgattc aatcagggtt tatttgactg tattgaactc tttttgtgtg tttgcagctc 5640
ataaaaagga tccaccatga acaagaacaa caccaagctg agcacccgcg ccctgccgag 5700
cttcatcgac tacttcaacg gcatctacgg cttcgccacc ggcatcaagg acatcatgaa 5760
catgatcttc aagaccgaca ccggcggcga cctgaccctg gacgagatcc tgaagaacca 5820
gcagctgctg aacgacatca gcggcaagct ggacggcgtg aacggcagcc tgaacgacct 5880
gatcgcccag ggcaacctga acaccgagct gagcaaggag atccttaaga tcgccaacga 5940
gcagaaccag gtgctgaacg acgtgaacaa caagctggac gccatcaaca ccatgctgcg 6000
cgtgtacctg ccgaagatca ccagcatgct gagcgacgtg atgaagcaga actacgccct 6060
gagcctgcag atcgagtacc tgagcaagca gctgcaggag atcagcgaca agctggacat 6120
catcaacgtg aacgtcctga tcaacagcac cctgaccgag atcaccccgg cctaccagcg 6180
catcaagtac gtgaacgaga agttcgaaga gctgaccttc gccaccgaga ccagcagcaa 6240
ggtgaagaag gacggcagcc cggccgacat cctggacgag ctgaccgagc tgaccgagct 6300
ggcgaagagc gtgaccaaga acgacgtgga cggcttcgag ttctacctga acaccttcca 6360
cgacgtgatg gtgggcaaca acctgttcgg ccgcagcgcc ctgaagaccg ccagcgagct 6420
gatcaccaag gagaacgtga agaccagcgg cagcgaggtg ggcaacgtgt acaacttcct 6480
gatcgtgctg accgccctgc aggcccaggc cttcctgacc ctgaccacct gtcgcaagct 6540
gctgggcctg gccgacatcg actacaccag catcatgaac gagcacttga acaaggagaa 6600
ggaggagttc cgcgtgaaca tcctgccgac cctgagcaac accttcagca acccgaacta 6660
cgccaaggtg aagggcagcg acgaggacgc caagatgatc gtggaggcta agccgggcca 6720
cgcgttgatc ggcttcgaga tcagcaacga cagcatcacc gtgctgaagg tgtacgaggc 6780
caagctgaag cagaactacc aggtggacaa ggacagcttg agcgaggtga tctacggcga 6840
catggacaag ctgctgtgtc cggaccagag cgagcaaatc tactacacca acaacatcgt 6900
gttcccgaac gagtacgtga tcaccaagat cgacttcacc aagaagatga agaccctgcg 6960
ctacgaggtg accgccaact tctacgacag cagcaccggc gagatcgacc tgaacaagaa 7020
gaaggtggag agcagcgagg ccgagtaccg caccctgagc gcgaacgacg acggcgtcta 7080
catgccactg ggcgtgatca gcgagacctt cctgaccccg atcaacggct ttggcctgca 7140
ggccgacgag aacagccgcc tgatcaccct gacctgtaag agctacctgc gcgagctgct 7200
gctagccacc gacctgagca acaaggagac caagctgatc gtgccaccga gcggcttcat 7260
cagcaacatc gtggagaacg gcagcatcga ggaggacaac ctggagccgt ggaaggccaa 7320
caacaagaac gcctacgtgg accacaccgg cggcgtgaac ggcaccaagg ccctgtacgt 7380
gcacaaggac ggcggcatca gccagttcat cggcgacaag ctgaagccga agaccgagta 7440
cgtgatccag tacaccgtga agggcaagcc atcgattcac ctgaaggacg agaacaccgg 7500
ctacatccac tacgaggaca ccaacaacaa cctggaggac taccagacca tcaacaagcg 7560
cttcaccacc ggcaccgacc tgaagggcgt gtacctgatc ctgaagagcc agaacggcga 7620
cgaggcctgg ggcgacaact tcatcatcct ggagatcagc ccgagcgaga agctgctgag 7680
cccggagctg atcaacacca acaactggac cagcaccggc agcaccaaca tcagcggcaa 7740
caccctgacc ctgtaccagg gcggccgcgg catcctgaag cagaacctgc agctggacag 7800
cttcagcacc taccgcgtgt acttcagcgt gagcggcgac gccaacgtgc gcatccgcaa 7860
ctcccgcgag gtgctgttcg agaagaggta catgagcggc gccaaggacg tgagcgagat 7920
gttcaccacc aagttcgaga aggacaactt ctacatcgag ctgagccagg gcaacaacct 7980
gtacggcggc ccgatcgtgc acttctacga cgtgagcatc aagtaggagc tctagatccc 8040
cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg 8100
ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta 8160
acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat 8220
acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg 8280
cggtgtcatc tatgttacta gatccctagg ggtccgatgt gagacttttc aacaaagggt 8340
aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta ttgtgaagat 8400
agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa aggccatcgt 8460
tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt 8520
ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atggtccgat 8580
tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat 8640
ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg 8700
cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc 8760
cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt 8820
ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca 8880
agacccttcc tctatataag gaagttcatt tcatttggag aggccatgga caacaaccca 8940
aacatcaacg aatgcattcc atacaactgc ttgagtaacc cagaagttga agtacttggt 9000
ggagaacgca ttgaaaccgg ttacactccc atcgacatct ccttgtcctt gacacagttt 9060
ctgctcagcg agttcgtgcc aggtgctggg ttcgttctcg gactagttga catcatctgg 9120
ggtatctttg gtccatctca atgggatgca ttcctggtgc aaattgagca gttgatcaac 9180
cagaggatcg aagagttcgc caggaaccag gccatctcta ggttggaagg attgagcaat 9240
ctctaccaaa tctatgcaga gagcttcaga gagtgggaag ccgatcctac taacccagct 9300
ctccgcgagg aaatgcgtat tcaattcaac gacatgaaca gcgccttgac cacagctatc 9360
ccattgttcg cagtccagaa ctaccaagtt cctctcttgt ccgtgtacgt tcaagcagct 9420
aatcttcacc tcagcgtgct tcgagacgtt agcgtgtttg ggcaaaggtg gggattcgat 9480
gctgcaacca tcaatagccg ttacaacgac cttactaggc tgattggaaa ctacaccgac 9540
cacgctgttc gttggtacaa cactggcttg gagcgtgtct ggggtcctga ttctagagat 9600
tggattagat acaaccagtt caggagagaa ttgaccctca cagttttgga cattgtgtct 9660
ctcttcccga actatgactc cagaacctac cctatccgta cagtgtccca acttaccaga 9720
gaaatctata ctaacccagt tcttgagaac ttcgacggta gcttccgtgg ttctgcccaa 9780
ggtatcgaag gctccatcag gagcccacac ttgatggaca tcttgaacag cataactatc 9840
tacaccgatg ctcacagagg agagtattac tggtctggac accagatcat ggcctctcca 9900
gttggattca gcgggcccga gtttaccttt cctctctatg gaactatggg aaacgccgct 9960
ccacaacaac gtatcgttgc tcaactaggt cagggtgtct acagaacctt gtcttccacc 10020
ttgtacagaa gacccttcaa tatcggtatc aacaaccagc aactttccgt tcttgacgga 10080
acagagttcg cctatggaac ctcttctaac ttgccatccg ctgtttacag aaagagcgga 10140
accgttgatt ccttggacga aatcccacca cagaacaaca atgtgccacc caggcaagga 10200
ttctcccaca ggttgagcca cgtgtccatg ttccgttccg gattcagcaa cagttccgtg 10260
agcatcatca gagctcctat gttctcttgg atacatcgta gtgctgagtt caacaacatc 10320
atcgcatccg atagtattac tcaaatccct gcagtgaagg gaaactttct cttcaacggt 10380
tctgtcattt caggaccagg attcactggt ggagacctcg ttagactcaa cagcagtgga 10440
aataacattc agaatagagg gtatattgaa gttccaattc acttcccatc cacatctacc 10500
agatatagag ttcgtgtgag gtatgcttct gtgaccccta ttcacctcaa cgttaattgg 10560
ggtaattcat ccatcttctc caatacagtt ccagctacag ctacctcctt ggataatctc 10620
caatccagcg atttcggtta ctttgaaagt gccaatgctt ttacatcttc actcggtaac 10680
atcgtgggtg ttagaaactt tagtgggact gcaggagtga ttatcgacag attcgagttc 10740
attccagtta ctgcaacact cgaggctgag tacaaccttg agagagccca gaaggctgtg 10800
aacgccctct ttacctccac caatcagctt ggcttgaaaa ctaacgttac tgactatcac 10860
attgaccaag tgtccaactt ggtcacctac cttagcgatg agttctgcct cgacgagaag 10920
cgtgaactct ccgagaaagt taaacacgcc aagcgtctca gcgacgagag gaatctcttg 10980
caagactcca acttcaaaga catcaacagg cagccagaac gtggttgggg tggaagcacc 11040
gggatcacca tccaaggagg cgacgatgtg ttcaaggaga actacgtcac cctctccgga 11100
actttcgacg agtgctaccc tacctacttg taccagaaga tcgatgagtc caaactcaaa 11160
gccttcacca ggtatcaact tagaggctac atcgaagaca gccaagacct tgaaatctac 11220
tcgatcaggt acaatgccaa gcacgagacc gtgaatgtcc caggtactgg ttccctctgg 11280
ccactttctg cccaatctcc cattgggaag tgtggagagc ctaacagatg cgctccacac 11340
cttgagtgga atcctgactt ggactgctcc tgcagggatg gcgagaagtg tgcccaccat 11400
tctcatcact tctccttgga catcgatgtg ggatgtactg acctgaatga ggacctcgga 11460
gtctgggtca tcttcaagat caagacccaa gacggacacg caagacttgg caaccttgag 11520
tttctcgaag agaaaccatt ggtcggtgaa gctctcgctc gtgtgaagag agcagagaag 11580
aagtggaggg acaaacgtga gaaactcgaa tgggaaacta acatcgttta caaggaggcc 11640
aaagagtccg tggatgcttt gttcgtgaac tcccaatatg atcagttgca agccgacacc 11700
aacatcgcca tgatccacgc cgcagacaaa cgtgtgcaca gcattcgtga ggcttacttg 11760
cctgagttgt ccgtgatccc tggtgtgaac gctgccatct tcgaggaact tgagggacgt 11820
atctttaccg cattctcctt gtacgatgcc agaaacgtca tcaagaacgg tgacttcaac 11880
aatggcctca gctgctggaa tgtgaaaggt catgtggacg tggaggaaca gaacaatcag 11940
cgttccgtcc tggttgtgcc tgagtgggaa gctgaagtgt cccaagaggt tagagtctgt 12000
ccaggtagag gctacattct ccgtgtgacc gcttacaagg agggatacgg tgagggttgc 12060
gtgaccatcc acgagatcga gaacaacacc gacgagctta agttctccaa ctgcgtcgag 12120
gaagaaatct atcccaacaa caccgttact tgcaacgact acactgtgaa tcaggaagag 12180
tacggaggtg cctacactag ccgtaacaga ggttacaacg aagctccttc cgttcctgct 12240
gactatgcct ccgtgtacga ggagaaatcc tacacagatg gcagacgtga gaacccttgc 12300
gagttcaaca gaggttacag ggactacaca ccacttccag ttggctatgt taccaaggag 12360
cttgagtact ttcctgagac cgacaaagtg tggatcgaga tcggtgaaac cgagggaacc 12420
ttcatcgtgg acagcgtgga gcttctcttg atggaggaat aatacgtagt cctttgtctt 12480
caattttgag ggctttttac tgaataagta tgtagtacta aaatgtatgc tgtaatagct 12540
catagtgagc gaggaaagta tcgggctatt taactatgac ttgagctcca tctatgaata 12600
aataaatcag catatgatgc ttttgttttg tgtacttcaa ctgtctgctt agctaatttg 12660
atatggttgg cacttggcac gtataaatat gctgaagtaa tttactctga agctaaatta 12720
actagattag atgagtgtat tatatacaaa aggcattaaa tcagatacat cttagacaaa 12780
ttgtcacggt ctaccagaaa agaaattgca tttgtttttg ggtctttcag actgacaaga 12840
tcgatctgaa gtctaaacaa ttctaagagg tatcatgtag caatgtcctg ccacaatatt 12900
gaattgaccc gggagattgt cgtttcccgc cttcagttta aactatcagt gtttgacagg 12960
atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa 13020
agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc 13080
cctcgggagt gcttggcatt ccgtgcgata atgacttctg ttcaaccacc caaacgtcgg 13140
aaagcctgac gacggagcag cattccaaaa agatcccttg gctcgtctgg gtcggctaga 13200
aggtcgagtg ggctgctgtg gcttgatccc tcaacgcggt cgcggacgta gcgcagcgcc 13260
gaaaaatcct cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 13320
catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 13380
cgtgacacca cgatgcaaaa gtcccatgtg gatcactccg ttgccccgtc gctcaccgtg 13440
ttggggggaa ggtgcacatg gctcagttct caatggaaat tatctgccta accggctcag 13500
ttctgcgtag aaaccaacat gcaagctcca ccgggtatga tggtcaatat ggagaaaaag 13560
aaagagtaat taccaatttt ttttcaattc aaaaatgtag atgtccgcag cgttattata 13620
aaatgaaagt acattttgat aaaacgacaa attacgatcc gtcgtattta taggcgaaag 13680
caataaacaa attattctaa ttcggaaatc tttatttcga cgtgtctaca ttcacgtcca 13740
aatgggggct tagatgaggc gcgccggtac cgtcgacatt gatgcatgtt gtcaatcaat 13800
tggcaagtca taaaatgcat taaaaaatat tttcatactc aactacaaat ccatgagtat 13860
aactataatt ataaagcaat gattagaatc tgacaaggat tctggaaaat tacataaagg 13920
aaagttcata aatgtctaaa acacaagagg acatacttgt attcagtaac atttgcagct 13980
tttctaggtc tgaaaatata tttgttgcct agtgaataag cataatggta caactacaag 14040
tgttttactc ctcatattaa cttcggtcat tagaggccac gatttgacac atttttactc 14100
aaaacaaaat gtttgcatat ctcttataat ttcaaattca acacacaaca aataagagaa 14160
aaaacaaata atattaattt gagaatgaac aaaaggacca tatcattcat taactcttct 14220
ccatccattt ccatttcaca gttcgatagc gaaaaccgaa taaaaaacac agtaaattac 14280
aagcacaaca aatggtacaa gaaaaacagt tttcccaatg ccataatact caaactcagt 14340
aggattctgg tgtgtgcgca atgaaactga tgcattgaac ttgacgaacg ttgtcgaaac 14400
cgatgatacg aacgaaagct gaattcttat ttgccgacta ccttggtgat ctcgcctttc 14460
acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt 14520
ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc 14580
attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa 14640
atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc 14700
catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag 14760
agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag 14820
atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc 14880
tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg 14940
tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa 15000
gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc 15060
gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac 15120
aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat 15180
agttgagtcg atacttcggc gatcaccgct tcccccatgc acgccgtgga aacagaagac 15240
atgaccttaa gaggacgaag ctcagagcca attaacgtca tcccactctt cttcaatccc 15300
cacgacgacg aaatcggata agctcgtgga tgctgctgcg tcttcagaga aaccgataag 15360
ggagatttgc gttgactgga tttcgagaga ttggagataa gagatgggtt ctgcacacca 15420
ttgcagattc tgctaacttg cgccatggta gttctctggt caacaaatct gcaaaaaaaa 15480
gaacaatcaa atattgaatc aataaaaaca gtagacatat ataaccatta tgaaaatcat 15540
ctttaacaca aaacaatcat agtaaaaagc cgctatcgga gtcaaaccga tttctaccac 15600
aaaaattgtg gtgctctatc atctgagcta agcgagctaa attcaatcaa atcggcaatt 15660
tcaagagata taaagcaaac aatccgagaa ttacccttag aaaaagggaa cactggattt 15720
gttatgctac tccagaatta cccaatcaca attcaaatcg acatacatct taaatacgca 15780
aatcaacaag ttcactcgta tatagatcca ccaaccaaca atcaatcgaa caaattaaaa 15840
tctcaaaaca agatcgatca gaatgggaaa atagtttcat gaataaatta tcgtttagaa 15900
gcagtgaaac gttgatatta acaatcgata aaatctaaac aacagatcag aattttcaga 15960
acgatcaatc aacaaagagc atcgaatcca acaatcatcg aaataatatg agcaacgatc 16020
agcagacgaa aaagaaaaag aacgatctaa gggtacgtta acgcttacct cagagatatc 16080
gcagaaaagt gtgtagcggc tagggtcaat aagaacgata aatggaatca gcaacgagag 16140
tggttttatt tatagcgttg aatccgaaac caatgttagg gtttaacata aaataatatc 16200
attctttaaa ttaccattat gcccctgttt agccgtgaat actcaatacc tcttgggttg 16260
gttgtgtttt atgaatgttt agtactacat gttacaaaca tcatttaatc acacctttta 16320
attaaggtta ttcaagcaaa tggttataca tgttcgtctc ccaaaaattt cattgattgt 16380
ggaccatcaa tctatatgac actcttctct agagtaaata tttgatatct caatgaatgc 16440
agattatcaa gagccattta tatcatttat tcaaacacca aataaaataa tgaacaagta 16500
aagtatatat taacctcaaa atctcatgac aatggaaatg gtttattata acaaccatat 16560
gtgtcccatt ccacgagcaa cctccctcaa gagagaaact tccagatctc ttccttacaa 16620
acctttgagg atgggagttc cttcttggtt ttggcgatac caatttgaat aaagtgatat 16680
ggctcgtacc ttgttgattg aacccaatct ggaatgctgc taaatcctga gctcaagcta 16740
attcttttgt ggtcgtcact gcgttcgtca tacgcattag tgagtgggct gtcaggacag 16800
ctcttttcca cgttattttg ttccccactt gtactagagg aatctgcttt atctttgcaa 16860
taaaggcaaa gatgcttttg gtaggtgcgc ctaacaattc tgcaccattc cttttttgtc 16920
tggtccccac aagccagctg ctcgatgttg acaagattac tttcaaagat gcccactaac 16980
tttaagtctt cggtggatgt ctttttctga aacttactga ccatgatgca tgtgctggaa 17040
cagtagttta ctttgattga agattcttca ttgatctcct gtagcttttg gctaatggtt 17100
tggagactct gtaccctgac cttgttgagg ctttggactg agaattaagc tgctagattg 17160
tcgtttcccg ccttcagttt aaactatcag tgtttg 17196
<210> 10
<211> 18708
<212> DNA
<213> Artificial Sequence
<400> 10
tacggcttat caccctcaat caaatggtta agcggaagtg tcgaattgag aaatcaagtt 60
gattttggag aagaccgtta agccagatag gaaagattag agtttgtgac taaatgatgc 120
actgtgggcg taccgtacag cttataaagg acccataggt atgtctcctt atcgactaat 180
ttttggtaaa ccgtgtcatc ttcctgttga actagaacat aaagcgtttc gggaggtcaa 240
gcaatgcaac atggagttgg aagttgcagg taagtatcgt cagttatata ttcaggaact 300
tgaagaaatt caacgagaag cgtatgaaag tgcccaaatt tataaggata agatcaaggc 360
ataccatgat cagaagatat cctgtaaata attttcggta cgacataaag tattattcta 420
tgaccctaca ttaagaattt ttgcaggtaa gcttcaaact aagtggttaa ggcattttgt 480
tattactcac gtatttccac atggtacagt cgaggttaaa agcgaaaaat ctgcaaaaaa 540
tttcaaggtc aatggacagc gattaaagtc tttctacgaa aattttcaag tccacatggt 600
cgaggaatta gtcctcgaag agccggttaa ataaatttcc aggtcgtcga gctaatgacg 660
ttaaacaaaa gcgctatttg ggagacaacc caaatttatt ttcatttatt taattttgag 720
tttaaataag atttaggttg taaataaata aataaataaa tcaatgtact tttcatgtcc 780
gggaaatcta catggatcag caatgagtat gatggtcaat atggagaaaa agaaagagta 840
attaccaatt ttttttcaat tcaaaaatgt agatgtccgc agcgttatta taaaatgaaa 900
gtacattttg ataaaacgac aaattacgat ccgtcgtatt tataggcgaa agcaataaac 960
aaattattct aattcggaaa tctttatttc gacgtgtcta cattcacgtc caaatggggg 1020
cttagatgag gcgcgccggt accgtcgaca ttgatgcatg ttgtcaatca attggcaagt 1080
cataaaatgc attaaaaaat attttcatac tcaactacaa atccatgagt ataactataa 1140
ttataaagca atgattagaa tctgacaagg attctggaaa attacataaa ggaaagttca 1200
taaatgtcta aaacacaaga ggacatactt gtattcagta acatttgcag cttttctagg 1260
tctgaaaata tatttgttgc ctagtgaata agcataatgg tacaactaca agtgttttac 1320
tcctcatatt aacttcggtc attagaggcc acgatttgac acatttttac tcaaaacaaa 1380
atgtttgcat atctcttata atttcaaatt caacacacaa caaataagag aaaaaacaaa 1440
taatattaat ttgagaatga acaaaaggac catatcattc attaactctt ctccatccat 1500
ttccatttca cagttcgata gcgaaaaccg aataaaaaac acagtaaatt acaagcacaa 1560
caaatggtac aagaaaaaca gttttcccaa tgccataata ctcaaactca gtaggattct 1620
ggtgtgtgcg caatgaaact gatgcattga acttgacgaa cgttgtcgaa accgatgata 1680
cgaacgaaag ctgaattctc aggcagcctt cgtatcggag agttcgatct tcgcgcccag 1740
cccggccatc aggtccatga actccgggaa gctcgtggcg atcatcgtgg catcgtccac 1800
cgtgacaggg ttttccgaca cgaggcccat gacgaggaag ctcatggcga tgcggtgatc 1860
gagatgggtg gcgacggcgg cgcccgaggc gttgccgagc cccttgccgt cagggcggcc 1920
acgcacgacg agcgacgtct cgccctcatc gcaatccacg ccattgagct tgaggccatt 1980
ggcgacggcc gagaggcggt cgctttcctt gacgcggagt tcttccagac cgttcatcac 2040
ggtcgcccct tccgcgaagg cggcggcgac agcgagaatc ggatattcgt cgatcatcga 2100
aggcgcgcgg tcttccggca ccgtgacgcc cttcagcgtg gaggagcgaa cgcgcaggtc 2160
cgccacgtct tcgccgccgg caaggcgcgg gttgatgact tcgatgtcgg cgcccatttc 2220
ctgcagcgtc aggatgaggc cggtgcgggt ggggttcatc agcacgttga ggatggtgac 2280
gtcggagccc ggaacaagca gggccgcaac cagcgggaag gccgtcgagg acgggtcgcc 2340
cggcacgtcg atgacttggc cggtgagctt gccgcggcct tccaggcgga tggtgcgcac 2400
gccgtccgca tccgtctcga cggtaaggtt ggcgccaaag ccctgcagca tcttttccgt 2460
atgatcgcgc gtcatgatcg gctcgatgac cgtcgtgatg ccgggcgtgt tgaggccggc 2520
gagcagcacg gcggacttca cctgtgcgga ggccatcggc acgcggtagg tgatcggcgt 2580
cggcgtcttc ggcccgcgca aggtaacggg aagacggtca ccgtcttccg atttcacctg 2640
cacgcccatt tcgcgcagcg ggttcaacac gcggcccatc gggcgctttg tgagcgaggc 2700
gtcgccgatg aaggtgctgt cgaaatcgta gaccccgacg aggcccatcg tcaggcggca 2760
gcccgtggcg gcattgccga aatcgagcgg cgcctcaggc gccaggaggc cgccattgcc 2820
gacgccatcg atgatccagg tgtcgccttc cttacggatg cgggcgccca tcgcctgcat 2880
ggccttgccc gtattgatga cgtcctcgcc ttccagaagg ccggtgatgc gcgtttcacc 2940
gctcgcgaga ccgccgaaca tgaaggaccg gtgggagatc gacttgtcgc cgggaatgcg 3000
gacggttccg gaaaggccag aggatttgcg ggcggttgcg ggccggctgc ttgcaccgtg 3060
aagcatgcac gccgtggaaa cagaagacat gaccttaaga ggacgaagct cagagccaat 3120
taacgtcatc ccactcttct tcaatcccca cgacgacgaa atcggataag ctcgtggatg 3180
ctgctgcgtc ttcagagaaa ccgataaggg agatttgcgt tgactggatt tcgagagatt 3240
ggagataaga gatgggttct gcacaccatt gcagattctg ctaacttgcg ccatggtagt 3300
tctctggtca acaaatctgc aaaaaaaaga acaatcaaat attgaatcaa taaaaacagt 3360
agacatatat aaccattatg aaaatcatct ttaacacaaa acaatcatag taaaaagccg 3420
ctatcggagt caaaccgatt tctaccacaa aaattgtggt gctctatcat ctgagctaag 3480
cgagctaaat tcaatcaaat cggcaatttc aagagatata aagcaaacaa tccgagaatt 3540
acccttagaa aaagggaaca ctggatttgt tatgctactc cagaattacc caatcacaat 3600
tcaaatcgac atacatctta aatacgcaaa tcaacaagtt cactcgtata tagatccacc 3660
aaccaacaat caatcgaaca aattaaaatc tcaaaacaag atcgatcaga atgggaaaat 3720
agtttcatga ataaattatc gtttagaagc agtgaaacgt tgatattaac aatcgataaa 3780
atctaaacaa cagatcagaa ttttcagaac gatcaatcaa caaagagcat cgaatccaac 3840
aatcatcgaa ataatatgag caacgatcag cagacgaaaa agaaaaagaa cgatctaagg 3900
gtacgttaac gcttacctca gagatatcgc agaaaagtgt gtagcggcta gggtcaataa 3960
gaacgataaa tggaatcagc aacgagagtg gttttattta tagcgttgaa tccgaaacca 4020
atgttagggt ttaacataaa ataatatcat tctttaaatt accattatgc ccctgtttag 4080
ccgtgaatac tcaatacctc ttgggttggt tgtgttttat gaatgtttag tactacatgt 4140
tacaaacatc atttaatcac accttttaat taaggttatt caagcaaatg gttatacatg 4200
ttcgtctccc aaaaatttca ttgattgtgg accatcaatc tatatgacac tcttctctag 4260
agtaaatatt tgatatctca atgaatgcag attatcaaga gccatttata tcatttattc 4320
aaacaccaaa taaaataatg aacaagtaaa gtatatatta acctcaaaat ctcatgacaa 4380
tggaaatggt ttattataac aaccatatgt gtcccattcc acgagcaacc tccctcaaga 4440
gagaaacttc cagatctctt ccttacaaac ctttgaggat gggagttcct tcttggtttt 4500
ggcgatacca atttgaataa agtgatatgg ctcgtacctt gttgattgaa cccaatctgg 4560
aatgctgcta aatcctgagc tcaagctaat tcttttgtgg tcgtcactgc gttcgtcata 4620
cgcattagtg agtgggctgt caggacagct cttttccacg ttattttgtt ccccacttgt 4680
actagaggaa tctgctttat ctttgcaata aaggcaaaga tgcttttggt aggtgcgcct 4740
aacaattctg caccattcct tttttgtctg gtccccacaa gccagctgct cgatgttgac 4800
aagattactt tcaaagatgc ccactaactt taagtcttcg gtggatgtct ttttctgaaa 4860
cttactgacc atgatgcatg tgctggaaca gtagtttact ttgattgaag attcttcatt 4920
gatctcctgt agcttttggc taatggtttg gagactctgt accctgacct tgttgaggct 4980
ttggactgag aattaagctt gtcgacaaaa tttagaacga acttaattat gatctcaaat 5040
acattgatac atatctcatc tagatctagg ttatcattat gtaagaaagt tttgacgaat 5100
atggcacgac aaaatggcta gactcgatgt aattggtatc tcaactcaac attatactta 5160
taccaaacat tagttagaca aaatttaaac aactattttt tatgtatgca agagtcagca 5220
tatgtataat tgattcagaa tcgttttgac gagttcggat gtagtagtag ccattattta 5280
atgtacatac taatcgtgaa tagtgaatat gatgaaacat tgtatcttat tgtataaata 5340
tccataaaca catcatgaaa gacactttct ttcacggtct gaattaatta tgatacaatt 5400
ctaatagaaa acgaattaaa ttacgttgaa ttgtatgaaa tctaattgaa caagccaacc 5460
acgacgacga ctaacgttgc ctggattgac tcggtttaag ttaaccacta aaaaaacgga 5520
gctgtcatgt aacacgcgga tcgagcaggt cacagtcatg aagccatcaa agcaaaagaa 5580
ctaatccaag ggctgagatg attaattagt ttaaaaatta gttaacacga gggaaaaggc 5640
tgtctgacag ccaggtcacg ttatctttac ctgtggtcga aatgattcgt gtctgtcgat 5700
tttaattatt tttttgaaag gccgaaaata aagttgtaag agataaaccc gcctatataa 5760
attcatatat tttcctctcc gctttgaatt gtctcgttgt cctcctcact ttcatcagcc 5820
gttttgaatc tccggcgact tgacagagaa gaacaaggaa gaagactaag agagaaagta 5880
agagataatc caggagattc attctccgtt ttgaatcttc ctcaatctca tcttcttccg 5940
ctctttcttt ccaaggtaat aggaactttc tggatctact ttatttgctg gatctcgatc 6000
ttgttttctc aatttccttg agatctggaa ttcgtttaat ttggatctgt gaacctccac 6060
taaatctttt ggttttacta gaatcgatct aagttgaccg atcagttagc tcgattatag 6120
ctaccagaat ttggcttgac cttgatggag agatccatgt tcatgttacc tgggaaatga 6180
tttgtatatg tgaattgaaa tctgaactgt tgaagttaga ttgaatctga acactgtcaa 6240
tgttagattg aatctgaaca ctgtttaagg ttagatgaag tttgtgtata gattcttcga 6300
aactttagga tttgtagtgt cgtacgttga acagaaagct atttctgatt caatcagggt 6360
ttatttgact gtattgaact ctttttgtgt gtttgcagct cataaaaagg atccaccatg 6420
aacaagaaca acaccaagct gagcacccgc gccctgccga gcttcatcga ctacttcaac 6480
ggcatctacg gcttcgccac cggcatcaag gacatcatga acatgatctt caagaccgac 6540
accggcggcg acctgaccct ggacgagatc ctgaagaacc agcagctgct gaacgacatc 6600
agcggcaagc tggacggcgt gaacggcagc ctgaacgacc tgatcgccca gggcaacctg 6660
aacaccgagc tgagcaagga gatccttaag atcgccaacg agcagaacca ggtgctgaac 6720
gacgtgaaca acaagctgga cgccatcaac accatgctgc gcgtgtacct gccgaagatc 6780
accagcatgc tgagcgacgt gatgaagcag aactacgccc tgagcctgca gatcgagtac 6840
ctgagcaagc agctgcagga gatcagcgac aagctggaca tcatcaacgt gaacgtcctg 6900
atcaacagca ccctgaccga gatcaccccg gcctaccagc gcatcaagta cgtgaacgag 6960
aagttcgaag agctgacctt cgccaccgag accagcagca aggtgaagaa ggacggcagc 7020
ccggccgaca tcctggacga gctgaccgag ctgaccgagc tggcgaagag cgtgaccaag 7080
aacgacgtgg acggcttcga gttctacctg aacaccttcc acgacgtgat ggtgggcaac 7140
aacctgttcg gccgcagcgc cctgaagacc gccagcgagc tgatcaccaa ggagaacgtg 7200
aagaccagcg gcagcgaggt gggcaacgtg tacaacttcc tgatcgtgct gaccgccctg 7260
caggcccagg ccttcctgac cctgaccacc tgtcgcaagc tgctgggcct ggccgacatc 7320
gactacacca gcatcatgaa cgagcacttg aacaaggaga aggaggagtt ccgcgtgaac 7380
atcctgccga ccctgagcaa caccttcagc aacccgaact acgccaaggt gaagggcagc 7440
gacgaggacg ccaagatgat cgtggaggct aagccgggcc acgcgttgat cggcttcgag 7500
atcagcaacg acagcatcac cgtgctgaag gtgtacgagg ccaagctgaa gcagaactac 7560
caggtggaca aggacagctt gagcgaggtg atctacggcg acatggacaa gctgctgtgt 7620
ccggaccaga gcgagcaaat ctactacacc aacaacatcg tgttcccgaa cgagtacgtg 7680
atcaccaaga tcgacttcac caagaagatg aagaccctgc gctacgaggt gaccgccaac 7740
ttctacgaca gcagcaccgg cgagatcgac ctgaacaaga agaaggtgga gagcagcgag 7800
gccgagtacc gcaccctgag cgcgaacgac gacggcgtct acatgccact gggcgtgatc 7860
agcgagacct tcctgacccc gatcaacggc tttggcctgc aggccgacga gaacagccgc 7920
ctgatcaccc tgacctgtaa gagctacctg cgcgagctgc tgctagccac cgacctgagc 7980
aacaaggaga ccaagctgat cgtgccaccg agcggcttca tcagcaacat cgtggagaac 8040
ggcagcatcg aggaggacaa cctggagccg tggaaggcca acaacaagaa cgcctacgtg 8100
gaccacaccg gcggcgtgaa cggcaccaag gccctgtacg tgcacaagga cggcggcatc 8160
agccagttca tcggcgacaa gctgaagccg aagaccgagt acgtgatcca gtacaccgtg 8220
aagggcaagc catcgattca cctgaaggac gagaacaccg gctacatcca ctacgaggac 8280
accaacaaca acctggagga ctaccagacc atcaacaagc gcttcaccac cggcaccgac 8340
ctgaagggcg tgtacctgat cctgaagagc cagaacggcg acgaggcctg gggcgacaac 8400
ttcatcatcc tggagatcag cccgagcgag aagctgctga gcccggagct gatcaacacc 8460
aacaactgga ccagcaccgg cagcaccaac atcagcggca acaccctgac cctgtaccag 8520
ggcggccgcg gcatcctgaa gcagaacctg cagctggaca gcttcagcac ctaccgcgtg 8580
tacttcagcg tgagcggcga cgccaacgtg cgcatccgca actcccgcga ggtgctgttc 8640
gagaagaggt acatgagcgg cgccaaggac gtgagcgaga tgttcaccac caagttcgag 8700
aaggacaact tctacatcga gctgagccag ggcaacaacc tgtacggcgg cccgatcgtg 8760
cacttctacg acgtgagcat caagtaggag ctctagatcc ccgaatttcc ccgatcgttc 8820
aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat 8880
catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt 8940
atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga 9000
aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact 9060
agatccctag gggtccgatg tgagactttt caacaaaggg taatatccgg aaacctcctc 9120
ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc 9180
tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac 9240
agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca 9300
accacgtctt caaagcaagt ggattgatgt gatggtccga ttgagacttt tcaacaaagg 9360
gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag 9420
atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc 9480
gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc 9540
gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc 9600
actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa 9660
ggaagttcat ttcatttgga gaggccatgg acaacaaccc aaacatcaac gaatgcattc 9720
catacaactg cttgagtaac ccagaagttg aagtacttgg tggagaacgc attgaaaccg 9780
gttacactcc catcgacatc tccttgtcct tgacacagtt tctgctcagc gagttcgtgc 9840
caggtgctgg gttcgttctc ggactagttg acatcatctg gggtatcttt ggtccatctc 9900
aatgggatgc attcctggtg caaattgagc agttgatcaa ccagaggatc gaagagttcg 9960
ccaggaacca ggccatctct aggttggaag gattgagcaa tctctaccaa atctatgcag 10020
agagcttcag agagtgggaa gccgatccta ctaacccagc tctccgcgag gaaatgcgta 10080
ttcaattcaa cgacatgaac agcgccttga ccacagctat cccattgttc gcagtccaga 10140
actaccaagt tcctctcttg tccgtgtacg ttcaagcagc taatcttcac ctcagcgtgc 10200
ttcgagacgt tagcgtgttt gggcaaaggt ggggattcga tgctgcaacc atcaatagcc 10260
gttacaacga ccttactagg ctgattggaa actacaccga ccacgctgtt cgttggtaca 10320
acactggctt ggagcgtgtc tggggtcctg attctagaga ttggattaga tacaaccagt 10380
tcaggagaga attgaccctc acagttttgg acattgtgtc tctcttcccg aactatgact 10440
ccagaaccta ccctatccgt acagtgtccc aacttaccag agaaatctat actaacccag 10500
ttcttgagaa cttcgacggt agcttccgtg gttctgccca aggtatcgaa ggctccatca 10560
ggagcccaca cttgatggac atcttgaaca gcataactat ctacaccgat gctcacagag 10620
gagagtatta ctggtctgga caccagatca tggcctctcc agttggattc agcgggcccg 10680
agtttacctt tcctctctat ggaactatgg gaaacgccgc tccacaacaa cgtatcgttg 10740
ctcaactagg tcagggtgtc tacagaacct tgtcttccac cttgtacaga agacccttca 10800
atatcggtat caacaaccag caactttccg ttcttgacgg aacagagttc gcctatggaa 10860
cctcttctaa cttgccatcc gctgtttaca gaaagagcgg aaccgttgat tccttggacg 10920
aaatcccacc acagaacaac aatgtgccac ccaggcaagg attctcccac aggttgagcc 10980
acgtgtccat gttccgttcc ggattcagca acagttccgt gagcatcatc agagctccta 11040
tgttctcttg gatacatcgt agtgctgagt tcaacaacat catcgcatcc gatagtatta 11100
ctcaaatccc tgcagtgaag ggaaactttc tcttcaacgg ttctgtcatt tcaggaccag 11160
gattcactgg tggagacctc gttagactca acagcagtgg aaataacatt cagaatagag 11220
ggtatattga agttccaatt cacttcccat ccacatctac cagatataga gttcgtgtga 11280
ggtatgcttc tgtgacccct attcacctca acgttaattg gggtaattca tccatcttct 11340
ccaatacagt tccagctaca gctacctcct tggataatct ccaatccagc gatttcggtt 11400
actttgaaag tgccaatgct tttacatctt cactcggtaa catcgtgggt gttagaaact 11460
ttagtgggac tgcaggagtg attatcgaca gattcgagtt cattccagtt actgcaacac 11520
tcgaggctga gtacaacctt gagagagccc agaaggctgt gaacgccctc tttacctcca 11580
ccaatcagct tggcttgaaa actaacgtta ctgactatca cattgaccaa gtgtccaact 11640
tggtcaccta ccttagcgat gagttctgcc tcgacgagaa gcgtgaactc tccgagaaag 11700
ttaaacacgc caagcgtctc agcgacgaga ggaatctctt gcaagactcc aacttcaaag 11760
acatcaacag gcagccagaa cgtggttggg gtggaagcac cgggatcacc atccaaggag 11820
gcgacgatgt gttcaaggag aactacgtca ccctctccgg aactttcgac gagtgctacc 11880
ctacctactt gtaccagaag atcgatgagt ccaaactcaa agccttcacc aggtatcaac 11940
ttagaggcta catcgaagac agccaagacc ttgaaatcta ctcgatcagg tacaatgcca 12000
agcacgagac cgtgaatgtc ccaggtactg gttccctctg gccactttct gcccaatctc 12060
ccattgggaa gtgtggagag cctaacagat gcgctccaca ccttgagtgg aatcctgact 12120
tggactgctc ctgcagggat ggcgagaagt gtgcccacca ttctcatcac ttctccttgg 12180
acatcgatgt gggatgtact gacctgaatg aggacctcgg agtctgggtc atcttcaaga 12240
tcaagaccca agacggacac gcaagacttg gcaaccttga gtttctcgaa gagaaaccat 12300
tggtcggtga agctctcgct cgtgtgaaga gagcagagaa gaagtggagg gacaaacgtg 12360
agaaactcga atgggaaact aacatcgttt acaaggaggc caaagagtcc gtggatgctt 12420
tgttcgtgaa ctcccaatat gatcagttgc aagccgacac caacatcgcc atgatccacg 12480
ccgcagacaa acgtgtgcac agcattcgtg aggcttactt gcctgagttg tccgtgatcc 12540
ctggtgtgaa cgctgccatc ttcgaggaac ttgagggacg tatctttacc gcattctcct 12600
tgtacgatgc cagaaacgtc atcaagaacg gtgacttcaa caatggcctc agctgctgga 12660
atgtgaaagg tcatgtggac gtggaggaac agaacaatca gcgttccgtc ctggttgtgc 12720
ctgagtggga agctgaagtg tcccaagagg ttagagtctg tccaggtaga ggctacattc 12780
tccgtgtgac cgcttacaag gagggatacg gtgagggttg cgtgaccatc cacgagatcg 12840
agaacaacac cgacgagctt aagttctcca actgcgtcga ggaagaaatc tatcccaaca 12900
acaccgttac ttgcaacgac tacactgtga atcaggaaga gtacggaggt gcctacacta 12960
gccgtaacag aggttacaac gaagctcctt ccgttcctgc tgactatgcc tccgtgtacg 13020
aggagaaatc ctacacagat ggcagacgtg agaacccttg cgagttcaac agaggttaca 13080
gggactacac accacttcca gttggctatg ttaccaagga gcttgagtac tttcctgaga 13140
ccgacaaagt gtggatcgag atcggtgaaa ccgagggaac cttcatcgtg gacagcgtgg 13200
agcttctctt gatggaggaa taatacgtag tcctttgtct tcaattttga gggcttttta 13260
ctgaataagt atgtagtact aaaatgtatg ctgtaatagc tcatagtgag cgaggaaagt 13320
atcgggctat ttaactatga cttgagctcc atctatgaat aaataaatca gcatatgatg 13380
cttttgtttt gtgtacttca actgtctgct tagctaattt gatatggttg gcacttggca 13440
cgtataaata tgctgaagta atttactctg aagctaaatt aactagatta gatgagtgta 13500
ttatatacaa aaggcattaa atcagataca tcttagacaa attgtcacgg tctaccagaa 13560
aagaaattgc atttgttttt gggtctttca gactgacaag atcgatctga agtctaaaca 13620
attctaagag gtatcatgta gcaatgtcct gccacaatat tgaattgacc cgggagattg 13680
tcgtttcccg ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc 13740
taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat 13800
ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggag tgcttggcat 13860
tccgtgcgat aatgacttct gttcaaccac ccaaacgtcg gaaagcctga cgacggagca 13920
gcattccaaa aagatccctt ggctcgtctg ggtcggctag aaggtcgagt gggctgctgt 13980
ggcttgatcc ctcaacgcgg tcgcggacgt agcgcagcgc cgaaaaatcc tcgatcggag 14040
gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 14100
gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcaaa 14160
agtcccatgt ggatcactcc gttgccccgt cgctcaccgt gttgggggga aggtgcacat 14220
ggctcagttc tcaatggaaa ttatctgcct aaccggctca gttctgcgta gaaaccaaca 14280
tgcaagctcc accgggtatg atggtcaata tggagaaaaa gaaagagtaa ttaccaattt 14340
tttttcaatt caaaaatgta gatgtccgca gcgttattat aaaatgaaag tacattttga 14400
taaaacgaca aattacgatc cgtcgtattt ataggcgaaa gcaataaaca aattattcta 14460
attcggaaat ctttatttcg acgtgtctac attcacgtcc aaatgggggc ttagatgagg 14520
cgcgccggta ccgtcgacat tgatgcatgt tgtcaatcaa ttggcaagtc ataaaatgca 14580
ttaaaaaata ttttcatact caactacaaa tccatgagta taactataat tataaagcaa 14640
tgattagaat ctgacaagga ttctggaaaa ttacataaag gaaagttcat aaatgtctaa 14700
aacacaagag gacatacttg tattcagtaa catttgcagc ttttctaggt ctgaaaatat 14760
atttgttgcc tagtgaataa gcataatggt acaactacaa gtgttttact cctcatatta 14820
acttcggtca ttagaggcca cgatttgaca catttttact caaaacaaaa tgtttgcata 14880
tctcttataa tttcaaattc aacacacaac aaataagaga aaaaacaaat aatattaatt 14940
tgagaatgaa caaaaggacc atatcattca ttaactcttc tccatccatt tccatttcac 15000
agttcgatag cgaaaaccga ataaaaaaca cagtaaatta caagcacaac aaatggtaca 15060
agaaaaacag ttttcccaat gccataatac tcaaactcag taggattctg gtgtgtgcgc 15120
aatgaaactg atgcattgaa cttgacgaac gttgtcgaaa ccgatgatac gaacgaaagc 15180
tgaattctta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt 15240
ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag 15300
cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga 15360
catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca 15420
ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat 15480
ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac 15540
ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga 15600
tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca 15660
gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt 15720
ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga 15780
tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat 15840
cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg 15900
tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg 15960
cgatcaccgc ttcccccatg cacgccgtgg aaacagaaga catgacctta agaggacgaa 16020
gctcagagcc aattaacgtc atcccactct tcttcaatcc ccacgacgac gaaatcggat 16080
aagctcgtgg atgctgctgc gtcttcagag aaaccgataa gggagatttg cgttgactgg 16140
atttcgagag attggagata agagatgggt tctgcacacc attgcagatt ctgctaactt 16200
gcgccatggt agttctctgg tcaacaaatc tgcaaaaaaa agaacaatca aatattgaat 16260
caataaaaac agtagacata tataaccatt atgaaaatca tctttaacac aaaacaatca 16320
tagtaaaaag ccgctatcgg agtcaaaccg atttctacca caaaaattgt ggtgctctat 16380
catctgagct aagcgagcta aattcaatca aatcggcaat ttcaagagat ataaagcaaa 16440
caatccgaga attaccctta gaaaaaggga acactggatt tgttatgcta ctccagaatt 16500
acccaatcac aattcaaatc gacatacatc ttaaatacgc aaatcaacaa gttcactcgt 16560
atatagatcc accaaccaac aatcaatcga acaaattaaa atctcaaaac aagatcgatc 16620
agaatgggaa aatagtttca tgaataaatt atcgtttaga agcagtgaaa cgttgatatt 16680
aacaatcgat aaaatctaaa caacagatca gaattttcag aacgatcaat caacaaagag 16740
catcgaatcc aacaatcatc gaaataatat gagcaacgat cagcagacga aaaagaaaaa 16800
gaacgatcta agggtacgtt aacgcttacc tcagagatat cgcagaaaag tgtgtagcgg 16860
ctagggtcaa taagaacgat aaatggaatc agcaacgaga gtggttttat ttatagcgtt 16920
gaatccgaaa ccaatgttag ggtttaacat aaaataatat cattctttaa attaccatta 16980
tgcccctgtt tagccgtgaa tactcaatac ctcttgggtt ggttgtgttt tatgaatgtt 17040
tagtactaca tgttacaaac atcatttaat cacacctttt aattaaggtt attcaagcaa 17100
atggttatac atgttcgtct cccaaaaatt tcattgattg tggaccatca atctatatga 17160
cactcttctc tagagtaaat atttgatatc tcaatgaatg cagattatca agagccattt 17220
atatcattta ttcaaacacc aaataaaata atgaacaagt aaagtatata ttaacctcaa 17280
aatctcatga caatggaaat ggtttattat aacaaccata tgtgtcccat tccacgagca 17340
acctccctca agagagaaac ttccagatct cttccttaca aacctttgag gatgggagtt 17400
ccttcttggt tttggcgata ccaatttgaa taaagtgata tggctcgtac cttgttgatt 17460
gaacccaatc tggaatgctg ctaaatcctg agctcaagct aattcttttg tggtcgtcac 17520
tgcgttcgtc atacgcatta gtgagtgggc tgtcaggaca gctcttttcc acgttatttt 17580
gttccccact tgtactagag gaatctgctt tatctttgca ataaaggcaa agatgctttt 17640
ggtaggtgcg cctaacaatt ctgcaccatt ccttttttgt ctggtcccca caagccagct 17700
gctcgatgtt gacaagatta ctttcaaaga tgcccactaa ctttaagtct tcggtggatg 17760
tctttttctg aaacttactg accatgatgc atgtgctgga acagtagttt actttgattg 17820
aagattcttc attgatctcc tgtagctttt ggctaatggt ttggagactc tgtaccctga 17880
ccttgttgag gctttggact gagaattaag ctgctagatt gtcgtttccc gccttcagtt 17940
taaactatca gtgtttgcaa ttatttacct acaaataaaa aaaccaacat ccctaacctt 18000
aaaaccccaa aacccaaatc caatacccac ccaccaacct aatacctacc ccaaatccat 18060
ttgtccaaac atcaaatgac ctatacaaaa aaaaccccaa tacccaaaac accaactagc 18120
ccaatatggc cccaaaaaat gaaaaggaag gaaattgggc cgaacgacct agctggccca 18180
aagaaaaaaa aagaacaaga aatggacaag aggcccaaac tgaccacaag gaaaaaaaaa 18240
gagaaaagag aaaaaaagaa aacgaaaaaa taaaacaccc aaaatcttta aaaccctaaa 18300
cccctttttt cttctccatc tttttcaaaa caaaacccta accccacact acaccgctgc 18360
cacctcaacc atcgttagag tcgtcgtcga gtgttgtcac caccatcatc gtccacattt 18420
gtcgtcgagc gctcacgagt caccatggtc tcgtgagcca ccattgtcga acgtcatttt 18480
tcagagtatc gttgctcacc ccattgccct atttcatcat cgtcgaactt agacaccacc 18540
acgttgacgc ggcatcgccg ttttacacca tcaccaacac gtcgtcgctc cgtccttcct 18600
caaacaccat ctcatctatt tcttcttttt ctccataatc accaaacacc caaaccaccc 18660
atcctaccac cttaaccgac gccatacctg aaacccccac cgtcaccc 18708
<210> 11
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 11
ccagaatcct tgtcagatt 19
<210> 12
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 12
tacggcttat caccctca 18
<210> 13
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 13
ataaaggcaa agatgctttt ggtaggtg 28
<210> 14
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 14
tttccttgtg gtcagttt 18
<210> 15
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 15
ggcagccttc gtatcggag 19
<210> 16
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 16
ctcgtgtcgg aaaaccctgt 20
<210> 17
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 17
gtccatgaac tccgggaagc tc 22
<210> 18
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 18
ctgaagtgtc ccaagaggtt agagtc 26
<210> 20
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 20
gttgttctcg atctcgtgga tg 22
<210> 21
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 21
ccgcttacaa ggagggatac ggtgagg 27
<210> 21
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 21
tccactacga ggacaccaac aac 23
<210> 22
<211> 21
<212> DNA
<213> Artificial Sequence
<400> 22
ccgttctggc tcttcaggat c 21
<210> 23
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 23
tggaggacta ccagaccatc aacaagcg 28
<210> 24
<211> 19
<212> DNA
<213> Artificial Sequence
<400> 24
agaatggcag cgcaatgac 19
<210> 25
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 25
cagcaagata gccagatcaa tgtc 24
<210> 26
<211> 26
<212> DNA
<213> Artificial Sequence
<400> 26
cttgcaggta tcttcgagcc agccac 26
<210> 27
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 27
aacttactga ccatgatgca tgtgctgg 28
<210> 28
<211> 43
<212> DNA
<213> Artificial Sequence
<400> 28
acgatggact ccagtcccag ctgctcgatg ttgacaagat tac 43

Claims (12)

1.一种棉花转化事件KJC003的特征基因序列,其特征在于,所述特征基因序列包含转基因表达盒,所述转基因表达盒的核苷酸序列为SEQ ID NO:9;所述特征序列位于四倍体棉花的D基因组第8组染色体上;所述特征基因序列还包含5'端连接序列,所述5'连接序列选自SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5或SEQ ID NO:7中的任一个;所述特征基因序列还包含3'端连接序列,所述3'端连接序列选自SEQ ID NO:2、SEQ ID NO:4、SEQ ID NO:6和SEQ ID NO:8中的任一个。
2.如权利要求1所述的棉花转化事件KJC003的特征基因序列,其特征在于,所述特征基因序列为核苷酸序列为SEQ ID NO:10的基因序列。
3.如权利要求1所述的棉花转化事件KJC003的特征基因序列的用途,其特征在于,该特征基因序列可以有效的提高棉花对草甘膦的耐受性。
4.如权利要求1所述的棉花转化事件KJC003的特征基因序列的用途,其特征在于,该特征基因序列可以有效的提高棉花对害虫侵染的抵抗。
5.如权利要求4所述的用途,其特征在于,所述害虫是鳞翅目的害虫。
6.如权利要求5所述的用途,其特征在于,所述鳞翅目的害虫是小地老虎或者棉铃虫。
7.用于检测权利要求1或2所述的棉花转化事件KJC003的特征基因序列的引物对,其特征在于,所述引物对包含特异性识别侧翼序列的第一引物对和第二引物对,所述第一引物对由SEQ ID NO:11和SEQ ID NO:12组成;所述第二引物对由SEQ ID NO:13和SEQ ID NO:14组成。
8.一种鉴定棉花生物样品中棉花转化事件KJC003的特征基因序列的方法,其特征在于,所述方法包括:(a)从待鉴定的棉花生物样品提取DNA样品;(b)以提取的DNA样品为模板,使用权利要求7所述的引物对进行PCR扩增;(c)检测PCR扩增产物的长度与棉花转化事件KJC003中的理论核苷酸序列长度比较。
9.一种产生对草甘膦除草剂耐受和抵抗鳞翅目害虫侵染的棉花植物的方法,其特征在于,包括向所述植物的基因组中引入权利要求1所述的棉花转化事件KJC003的特征基因序列。
10.如权利要求9所述的方法,其特征在于,所述方法包含:
a)将包含权利要求1所述的棉花转化事件KJC003的特征基因序列的棉花植物与缺乏事件KJC003特征基因序列的棉花植物杂交来产生子代植物;
b)选择包含权利要求1所述的棉花转化事件KJC003特征基因序列的第一子代植物。
11.如权利要求10所述的方法,其特征在于,所述方法进一步包括使所述第一子代植物自交来产生第二代子代植物,并选择对于所述棉花转化事件KJC003特征基因序列是纯合的植物。
12.权利要求1-4中任一项所述的棉花转化事件KJC003的特征基因序列用于提高棉花抗草甘膦性状、抗虫性状,进行棉花育种或用作分子标记的用途。
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