辣椒上CMV亚组ⅠB CP基因克隆与原核表达  

Cloning and Prokaryotic Expression of Coat Protein Gene of Cucumber Mosaic Virus subgroupⅠB Separated from Chilli Pepper in Xinjiang

作　　者：张强[1] 陈英杰[1] 向本春[1] 郑银英[1] 

机构地区：[1]石河子大学生命科学学院,新疆石河子832003

出　　处：《新疆农业科学》2014年第12期2257-2261,共5页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(31260420);国际科技合作与交流专项(20072072)

摘　　要：【目的】克隆辣椒上黄瓜花叶病毒外壳蛋白(CMV CP)基因,诱导CP蛋白表达,为下一步制备高效、特异的抗血清奠定基础。【方法】根据已知的新疆石河子辣椒上的CP基因序列设计引物,克隆CMV CP基因,经测序、NocⅠ/Bam HⅠ酶切鉴定,成功构建了原核表达载体p ET-CMV CP,转化大肠杆菌BL21 DE3,用终浓度为1 mmol/L IPTG诱导蛋白表达,SDS-PAGE检测目的蛋白表达。【结果】成功构建了新疆辣椒上CMV CP原核表达载体p ET-CMV CP,获得了与预期大小一致的30 k Da蛋白。【结论】新疆石河子辣椒上CMV CP在原核中获得了正确表达。[ Objective] The project aims to clone the CMV CP gene of chilli pepper in Shihezi and and induce the expression of CMV CP protein in the hope of laying a solid foundation for preparing a kind of antiserum with high efficiency and specificity. [ Method ] According to the known nueleotide sequence of CP gene separated from pepper in Shihezi, a pair of primers was designed, Sequencing and Noc I/Barn H I enzyme digestion were performed to ensure that the prokaryotic expression vector pET - CMV CP would be successfully constructed. The recombinant vector was transformed into BL21 DE3 host strain of E. coli. The expression of the objective protein was induced by IPTG at terminal concentration of l mmol/L and detected by SDS - PAGE. [ Result]The prokaryotie expression vector pET - CMV CP of CMV CP of pepper in Shihezi was sueeessfully constructed and expressed as a 30kDa protein, whose molecular weight was identical to the expected fusion protein. [ Conclusion]The CMV CP gene of pepper in Shihezi was correctly expressed in prokaryote.

关 键 词：辣椒 黄瓜花叶病毒 外壳蛋白 原核表达 

分 类 号：S641.3[农业科学—蔬菜学] S188[农业科学—园艺学]