甘蔗花叶病毒河南分离物HC-Pro基因的序列分析及其RNA沉默表达载体构建  

Sequence analysis and construction of RNA silencing expression vector of the Hc-Pro gene from Sugarcane mosaic virus-Henan isolate

作　　者：夏宗良[1] 武轲[1] 周朋[1] 吴建宇[1] 

机构地区：[1]河南农业大学生命科学学院,河南郑州450002

出　　处：《河南农业大学学报》2010年第2期191-195,共5页Journal of Henan Agricultural University

基　　金：河南省科技攻关重点项目(082102140003)

摘　　要：采用RT-PCR方法从河南地区甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)感染的玉米叶片中,克隆了辅助成分-蛋白酶基因(helper component-proteinase,HC-Pro).该基因长1 380 bp,编码460个氨基酸.序列比对表明,HC-Pro编码的氨基酸序列与北京、山东地区SCMV分离物的相似性分别达到97%和98%.而与以前数据库登录的河南分离物的相似性仅98%,说明河南地区SCMV可能发生了变异.选取长582 bp的HC-Pro片段,构建了表达发夹RNA的基因沉默双元表达载体pRNAi-HC-Pro(±),成功转化了农杆菌LBA4404.The complete helper component-proteinase （HC-Pro） gene was cloned by RT-PCR from diseased maize leaves infected by Henan isolate of Sugarcane mosaic virus （SCMV）. The cDNA of the HC-Pro contains a 1 380 bp open reading frame （ORF） and encodes 460 amino acids. Sequence alignment showed that the deduced amino acid sequence of the HC-Pro shared 97% and 98% similarities to homologues from SCMV Beijing isolate and Shandong isolate respectively. However, the Hc-Pro cloned by our group only shared 98% similarity to that of SCMV Henan isolate deposited in the Gen- bank, and this might indicate that the SCMV Henan isolate has undergone some genetic divesities. A 582 bp long fragment of the HC-Pro was selected to construct the binary expression vector pRNAi-HC- Pro （ ± ） with hairpin RNA structure, and the vector was successfully transformed into the agrobaeterium strain LBA4404.

关 键 词：甘蔗花叶病毒 玉米 辅助成分-蛋白酶 RNA干扰 

分 类 号：Q782[生物学—分子生物学] S513[农业科学—作物学]