番茄WRKY转录因子基因片段的克隆及序列分析  

Cloning and Sequence Analysis of WRKY Transcription Factor Gene Fragment in Tomato

作　　者：王丽芳[1,2] 杜希华[1] 于涌鲲[2] 赵福宽[2] 孙清鹏[2] 

机构地区：[1]山东师范大学生命科学学院,济南250014 [2]北京农学院生物技术系,北京102206

出　　处：《中国农学通报》2009年第23期70-73,共4页Chinese Agricultural Science Bulletin

基　　金：北京市属市管高等学校人才强教计划;北京市委组织部优秀人才培养资助项目(20042D0502108)

摘　　要：以番茄苗为实验材料,克隆出番茄WRKY转录因子的一个基因片段并对其核酸序列进行分析。根据笔者课题组已经克隆出的番茄特异的WRKY基因片段和番茄EST序列设计特异引物,利用RT-PCR技术获得番茄WRKY转录因子的一个基因片段并连接到pMD19-T载体中,转化DH5α,筛选阳性克隆,用双酶切分析法对其进行鉴定并进一步对核酸序列进行测序分析。结果表明,克隆获得的WRKY转录因子基因片段约为680 bp,用GenBank中的Blastn程序分析表明,其与CaWRKY(AY789641.1)、WIZZ(wizz mRNA)(AB028022.1)的相似性分别为86%和84%。WRKY转录因子在番茄中同样受JA诱导,这为克隆番茄WRKY基因全长,进而为番茄抗病育种奠定基础。In this study, tomato seedlings were used as gene fragment was cloned and sequenced. The specific experimental materials; a WRKY transcription factor primer was designed according to the cloned WRKY transcription factor gene fragment and tomato EST sequences; a gene fragment was amplified by RT-PCR. The fragment was inserted into the pMD19-T vector, and transformed into E. coli DH5ct.The recombinant plasmid was digested with Pst I and EcoR I, and sequenced. The result showed that the fragment was about 680 bp. Blastn analysis showed thatthe similarity with CaWRKY （AY789641.1） and WIZZ （wizz mRNA） （AB028022.1） were 86% and 84% respectively. WRKY transcription factor in tomato was induced by JA, which laid the foun- dation for cloning the full WRKY gene sequence in tomato and breeding disease-resistant tomato.

关 键 词：番茄 转录因子 WRKY 克隆 

分 类 号：S188[农业科学—农业基础科学]