美洲南瓜WRKY4基因克隆与序列分析  

Cloning and Sequence Analysis of WRKY4 Gene in Zucchini(Cucurbita pepo)

作　　者：徐岭贤[1] 赵福宽[1] 孙清鹏[1] 郝芮[1] 

机构地区：[1]北京农学院生物技术学院,北京102206

出　　处：《分子植物育种》2012年第6期701-706,共6页Molecular Plant Breeding

基　　金：北京市属高等学校人才强教计划资助项目(PHR200907136)资助

摘　　要：以美洲南瓜(Cucurbita pepo)幼叶为材料,采用Trizol方法进行总RNA的提取。根据GenBank上已提交的黄瓜WRKY4基因序列保守区域设计特异性引物,扩增得到1条长度为497bp的WRKY4转录因子基因片段,利用已知中间序列,通过RT-PCR和RACE(rapid amplification cDNA ends)等技术,克隆得到美洲南瓜WRKY4的cDNA全长序列,长度为1722bp,5'非翻译区为185bp,3'非翻译区为118bp,开放阅读框为1419bp,编码472个氨基酸,分子量约为51.40kD,等电点约为8.36,GenBank登录号为JX096811。用Blast和DNAMAN软件对其核苷酸序列进行分析,结果显示,美洲南瓜WRKY4与黄瓜WRKY4同源性达到87%。美洲南瓜WRKY4基因克隆及序列分析为以后研究此基因的功能奠定了基础。Total RNA was extracted from leaves of zucchini（Cucurbita pepo） byusing Trizol method.A cDNA fragment with 497 bp in length was amplified by specific primers designed based on the conserved domains of published WRKY4 sequences of cucumber in GenBank.Then full length of zucchini WRKY4 was obtained by RT-PCR and RACE that was deposited in GenBank with Accession No JX096811.The zucchini WRKY4 cDNA has 1 722 bp in length including 5＇ untranslated region of 185 bp,3＇ untranslated region of 118 bp with poly A,and an open reading frame of 1 419 bp encoding putative protein of 472 amino acids,the molecular weight of putative protein is 51.40 kD and its pI is 8.36.The nucleotide sequences were analyzed by using BLAST and DNAMAN software and the results showed that identity between zucchini WRKY4 and cucumber WRKY4 reached 87%.The cloning of WRKY4 gene from zucchini might provide the basis for further studying the role of WRKY4.

关 键 词：美洲南瓜 WRKY4 GenBank登录号JX096811 序列分析 

分 类 号：S642.1[农业科学—蔬菜学]