热带作物学报2009，Vol.30Issue(9)：1345-1350,6.
香蕉束顶病毒海口分离物NSP基因的原核表达
Prokaryotic Expression of NSP Gene of‘Haikou’Isolate of Banana Bunchy Top Virus
摘要
Abstract
The opening reading frame of NSP gene of banana bunchy top virus was cloned by polymerase chain reaction (PCR) using total DNA isolated from tender pseudostem and leaf of banana plant infected with the virus as a template, and directionally cloned to the downstream of 6 His tag by the Gateway technology. The construction of prokaryotic expressive vector, pDESTTM-17-NSP have been identified by the colony PCR and sequencing, and transformed into E.coli BL21(DE3) . Transformants were cultured and induced by IPTG, and then analyzed by SDS-PAGE and Western blot. The results showed that the fusion protein of approximately 20 ku was expressed steadily as an inclusion body and accorded with the predicted size. The expression of NSP gene was optimized under the conditions that the transformants were induced with 0.1 mmo1/1 IPTG at 25℃for 4 hours. This result might provide a foundation for preparation of the polyclonal antibody against NSP and further functional study of NSP.关键词
香蕉束顶病毒/Gateway重组技术/NSP/原核表达Key words
Banana bunchy top virus/gateway recombinant technology/NSP/prokaryotic expression.分类
农业科技引用本文复制引用
冯团诚,王健华,刘志昕..香蕉束顶病毒海口分离物NSP基因的原核表达[J].热带作物学报,2009,30(9):1345-1350,6.基金项目
国家科技支撑计划项目(No.2007BAD48801),教育部/海南省教育厅联合资助博士点基金项目(No.20050565002)资助. （No.2007BAD48801）
