动物医学进展2017，Vol.38Issue(4)：1-6,6.
猪圆环病毒2型ORF2基因的原核表达及反应原性分析
Prokaryotic Expression of ORF2 Gene Fragment of Porcine Circovirus Type 2 and the Reactinogenicity Analysis of the Expressed Protein
摘要
Abstract
This experiment was aimed to study the antigenicity of Cap protein of porcine circovirus type 2(PCV2).The gene was amplified from the DNA of PCV2-CAU0673 strain by PCR,whose product was approximately 702 bp.The product was cloned into pET30a(+)vector,PCR,digestion and sequencing were used to identify positive plasmid.The positive recombined plasmid was expressed by E.coli BL21(DE3)and was induced by IPTG.The recombinant protein was purified by Ni-NTA and refolding.The results of SDS-PAGE showed that the recombined protein was successfully expressed in E.coli BL21(DE3)with a relative molecular weight of 34 ku.The results of Western blot showed that this recombined protein was specifically reacted with PCV2 positive serum;no cross reacted with NA-PRRSV and PPV1 positive serum.The recombinant vector pET30a-PCV2-ORF2 was successfully constructed,and the recombined Cap protein with excellent reactinogenicity was successfully expressed in E.coli which laid a foundation for the diagnosis of PCV2.关键词
猪圆环病毒2型/ORF2基因/原核表达/反应原性Key words
Porcine circovirus type 2/ORF2 gene/prokaryotic expression/reactinogenicity分类
农业科技引用本文复制引用
欧云文,马小元,王俊,丁耀忠,张永光,张杰..猪圆环病毒2型ORF2基因的原核表达及反应原性分析[J].动物医学进展,2017,38(4):1-6,6.基金项目
国家国际合作项目 (2012DFG31890) （2012DFG31890）
国家自然科学基金项目(31072143) （31072143）
