绵羊MAVS基因的原核表达及多克隆抗体的制备  

PROKARYOTIC EXPRESSION AND POLYCLONAL ANTIBODY PREPARATION OF MAVS GENE IN OVIS ARIES

作　　者：董丹丹 刘腾[1] 缪秋红[1] 朱杰[1] 刘光清[1] DONG Dan-dan;LIU Teng;MIAO Qiu-hong;ZHU Jie;LIU Guang-qing(Shanghai Veterinary Research Institute,CAAS,Shanghai200241,China)

机构地区：[1]中国农业科学院上海兽医研究所

出　　处：《中国动物传染病学报》2019年第5期28-31,共4页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点研发计划项目(2016YFD0500108);国家自然科学基金项目(31502068);上海市科技兴农重点攻关项目(2016043)

摘　　要：通过RT-PCR方法从绵羊肺细胞(sheep lung cells,SLT)扩增获得MAVS(mitochondrial antiviral signaling protein)基因,并将其克隆至原核表达载体pET-32a,测序鉴定结果表明成功获得重组质粒pET-32a-MAVS。将其转化至感受肽细胞BL21中,经IPTG诱导获得重组蛋白。将纯化的MAVS蛋白免疫BALB/c雌鼠,制备抗MAVS的鼠源多克隆抗体。SDS-PAGE结果表明该抗体具有良好的反应原性。MAVS多克隆抗体的成功制备为其生物学功能研究奠定了基础。MAVS(mitochondrial antiviral signaling protein)gene was amplified from sheep lung cells(SLT)through RT-PCR and then was cloned into prokaryotic expression vector pET-32a.The recombinant plasmid was verified successfully by sequencing and named pET-32a-MAVS,which was transformed into BL21 and to obtain recombinant protein by IPTG induction.The purified MAVS protein was immunized to BALB/c female mice to obtain a polyclonal antibody.The result of SDS-PAGE proves that the antibody had a specifical immunogenicity.The successful preparation of MAVS polyclonal antibody has laid a foundation for the study of its biological function.

关 键 词：MAVS基因 原核表达 多克隆抗体 

分 类 号：S852.42[农业科学—基础兽医学]