香蕉条斑病毒CP区基因的克隆及其在大肠杆菌中的融合表达  

Cloning of Coat Protein Gene of Banana Streak Virus and its Fusion Expression in Escherichia coli

作　　者：邱世明[1] 庄军[1] 刘志昕[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所热带作物生物技术国家重点实验室,海南海口571101

出　　处：《安徽农业科学》2007年第23期7108-7109,共2页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金资助项目(30660100);海南省教育厅高等学校科研资助项目(Hjkj200505)

摘　　要：依据锌指结构在BSVCP上的可能位置合成了2条引物,通过PCR扩增得到长约640 bp的目的片段;将目的片段与质粒pET28 b(+)连接,构建了含BSVCP区基因的融合蛋白原核表达载体pET28b-BSVCP;克隆测序以确定其阅读编码框的正确性,然后用正确的重组质粒转化大肠杆菌Rosetta(DE3),并诱导表达;经IPTG诱导后,出现一特异的约30 kDa融合蛋白质表达条带。Banana streak disease caused by Banana streak virus （BSV） is now recognized as one of the most widely distributed viral diseases of plantains and banana. It is an important limiting factor to banana production. According to the location of zinc finger in the coat protein （CP） of BSV, two primers were designed and an about 650 bp DNA fragment was obtained by PCR. The PCR product, after digesting with restriction enzymes, was inserted into prokaryotic expression vector pET28b and the recombinant plasmid pET28b-BSVCP was transferred into E. coli Rosetta （DE3） after confirming the reading frame by sequencing. When pET28b-BSVCP was induced using IPTG, an additional 30 kDa protein band appeared by running SDS-PAGE. The efficient expression of BSV CP laid a firm foundation for further study on BSV CP and preparation viral antiserum for detecting BSV.

关 键 词：香蕉条斑病毒 外壳蛋白 原核表达 

分 类 号：Q78[生物学—分子生物学]