猪细小病毒自然弱毒N株非结构蛋白NS1基因原核表达  

Expression of non-structural protein NS1 gene in porcine parvovirus (PPV-N) strain

作　　者：苏乾莲[1] 李斌[1] 赵武[1] 梁家幸[1] 梁保忠[1] 何颖[1] 秦毅斌[1,2] 何苹萍[1,2] 

机构地区：[1]广西兽医研究所,南宁530001 [2]广西大学动物科学技术学院,南宁530005

出　　处：《广西农业科学》2010年第7期710-713,共4页Guangxi 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：为进一步探讨猪细小病毒自然弱毒N株(PPV-N株)的分子病原学机理,根据GenBank上已发表的PPV全基因组序列(登录号NC_001718)设计1对引物,通过PCR方法扩增获得PPV-N株NS1全长基因,将其克隆到pMD18-T载体上,进而构建原核表达重组质粒pET32a-NS1,并在BL21(DE3)pLysS中进行IPTG诱导表达。SDS-PAGE电泳显示NS1在大肠杆菌中得到成功表达,重组蛋白大小约为90.0 kDa;Western blotting分析表明,该蛋白能与PPV阳性血清发生特异性反应,证明该蛋白具有良好生物学活性。NS1在大肠杆菌中成功表达,且具有免疫原性,可作为PPV临床鉴别诊断抗原。The present experiment was conducted to study the molecular etiological mechanism of porcine parvovirus N strain.A pair of primer was designed from PPV genome sequences published in GenBank（Accession number NC-001718）.The whole genic sequence of NS1 in PPV-N strain was amplified by using PCR technology and cloned into pMD18-T vector.The recombinant of pET32a-NS1 was transformed into BL21（DE3） pLysS competent cell and induced to express by IPTG.SDS-PAGE results showed that the NS1 protein was successfully expressed in E.coli with a relative molecular weight of 90 kDa.The results of Western blotting showed that this protein was specifically reacted with porcine parvovirus positive serum,indicating the recombinant protein had biological activity.NS1 protein was expressed in E.coli and showed immunogenicity,which could be used as antigen in diagnostic assay for detection of antibodies of PPV.

关 键 词：猪细小病毒 自然弱毒N株 NS1 原核表达 

分 类 号：S858.28[农业科学—临床兽医学]