猪细小病毒自然弱毒N株VP2基因的克隆及原核表达  

Cloning and Prokaryotic Expression of VP2 Gene of Natural Attenuated Porcine Parvovirus N Strain

作　　者：李斌[1] 苏乾莲[1] 赵武[1] 梁家幸[1] 梁保忠[1] 何颖[1] 秦毅斌[1] 何苹萍[1] 

机构地区：[1]广西兽医研究所,广西南宁530001

出　　处：《西南农业学报》2011年第1期310-314,共5页Southwest China Journal of Agricultural Sciences

基　　金：广西自然科学基金项目(桂科自0728102);广西基本科研业务费专项项目(桂兽研专项09-4)

摘　　要：为了进一步研究猪细小病毒自然弱毒株(PPV-N株)的自然弱毒分子生物学机理,对PPV-N株VP2基因进行克隆、测序和原核表达研究。结果表明,成功构建PPV-N株VP2基因的克隆重组质粒pMD18-T-VP2及表达重组质粒pET32a-VP2,经SDS-PAGE及Western blotting鉴定,PPV-N株VP2基因在BL21(DE3)plysS菌中成功进行融合表达,表达出约85.4 KDa的VP2融合蛋白,且表达的VP2融合蛋白能与PPV阳性血清发生特异性反应。PPV-N株VP2基因的成功克隆及原核表达为今后研究PPV-N株的自然弱毒的分子生物学机理和研制PPV诊断抗原奠定了基础。In order to study the natural attenuated molecular biological mechanism of porcine parvovirus N strain,the VP2 gene of natural attenuated porcine parvovirus N strain（PPV-N strain） was cloned,sequenced and prokaryotic was expressed.The sequencing result showed that recombinant plasmid pMD18-T-VP2 and pET32a-VP2 of VP2 gene of PPV-N strain were constructed successfully.SDS-PAGE and Western blotting results showed that VP2 protein of PPV-N strain had fusion expression in the BL21（DE3）plysS,about 85.4 KDa VP2 fusion protein was expressed,and the VP2 fusion protein could react with pig serum containing antibody against PPV.The experimental results will provide foundation for study of natural attenuated molecular mechanism of PPV-N strain and developing diagnosable antigen for PPV.

关 键 词：猪细小病毒自然弱毒株(PPV-N株) VP2基因 克隆 原核表达 

分 类 号：S858.28[农业科学—临床兽医学]