兔补体C3d基因cDNA的克隆、鉴定及原核表达  

Cloning,indentification and expression of rabbit C3d cDNA

作　　者：魏后军[1] 王芳[1] 胡波[1] 范志宇[1] 薛家宾[1] 徐为中[1] 

机构地区：[1]江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《中国兽医学报》2013年第2期201-205,共5页Chinese Journal of Veterinary Science

基　　金：现代农业产业技术体系建设专项资金资助项目(CARS-44)

摘　　要：以兔肝脏组织中提取的总RNA为模板,通过巢式RT-PCR扩增出C3dcDNA,克隆测序后将其连接至表达载体pET-32a,转化入E.coli BL21,用IPTG诱导表达,并对表达产物进行SDS-PAGE和Western blot分析。SDS-PAGE分析表明,目的蛋白以包涵体形式存在并得到高效表达;Western blot进一步证实,表达的蛋白为C3d。本研究为开发利用兔C3d奠定了基础。The C3d gene from rabbit was amplified from total RNA by nested RT-PCR. The positive plasmid which contained C3d cDNA was determined by cloning and sequencing. After the fragments were cloned into pET-32a,the recombinant plasmid was transformed into E. coli BL21 ,and the expressed product was induced with IPTG. The expressed product was analyzed by SDS-PAGE and Western blot. The results showed that the fusion protein was expressed in a form of inclusion body. Western blot test showed that the expression protein was Cad. The research provides a foundation for development and utilization of rabbit C3d.

关 键 词：兔C3d 巢式RTPCR 原核表达 

分 类 号：S852.651[农业科学—基础兽医学] Q78[农业科学—兽医学]