甘蔗黄叶病毒外壳蛋白基因原核表达研究  

Sugarcane Yellow Leaf Virus Coat-Protein Gene Prokaryo Expression

作　　者：黄振瑞[1] 孟岩[1] 高三基[1] 许莉萍[1] 郭晋隆[1] 

机构地区：[1]福建农林大学/农业部甘蔗生理生态与遗传改良重点开放实验室

出　　处：《植物遗传资源学报》2007年第1期7-10,共4页Journal of Plant Genetic Resources

基　　金：农业部948计划资助项目(2006-G37);福建省自然科学基金资助项目(Z0516013)

摘　　要：本研究主要目的是构建ScYLV-CP基因的原核表达载体,表达ScYLV-CP蛋白。采用PCR扩增出CP基因的蛋白编码序列,克隆到中间载体中,再经双酶切、亚克隆到表达载体中,构建该基因的表达载体pET-29 a-CP,在大肠杆菌中用IPTG诱导表达,SDS-PAGE电泳。酶切鉴定表明表达载体内插入片段正确,在大肠杆菌中诱导后,出现一条分子量约为22 kDa蛋白带,与CP基因开放阅读框架的理论推算值21.719 kDa相符。研究表明甘蔗黄叶病毒外壳蛋白能在原核细胞中高效表达,为建立甘蔗黄叶病毒血清学快速检测技术奠定基础。The objective was to construct ScYLV-CP prokaryotic expression vector and express ScYLV-CP protein. PCR was used to obtain coding region of CP. Construction of a high-level protein expression vector pET-29a-CP was conducted by inserting the fragment of coding region of cp into protein expression vector pET-29a（＋）. Then the recombinant plasmid was transferred into E. coli to prepare protein. The result of endonucleases digesting shown that pET-29a-CP was constructed successfully. A new protein band of 22 kDa was observed by SDS-PAGE analysis after induction by ITPG, it is similar to the theoretic value of 21. 719 kDa according to the ORF of the CP gene. The conclusion was that sugarcane yellow leaf virus coat-protein could he expressed in E. coli expression system and purified initially.

关 键 词：甘蔗黄叶病毒 外壳蛋白基因 表达载体构建 原核表达 

分 类 号：S435.661[农业科学—农业昆虫与害虫防治]