摘要
A pair of primers containing BamHI and XhoI sites was designed to amplify n gene of 1149bp from the positive pMD18-TN plasmid of transmissible gastroenteritis virus n gene by PCR.PCR product was subcloned into multiple cloning sites of pET-30a containing 6 His·Tag.Recombinant plasmid pET-30a of n gene named pET30a-N was transfected into E.coli BL21 and the bacteria was induced with IPTG at 37℃.It was demonstrated by SDS-PAGE that recombinant N protein expressed in E.coli BL21 was soluble protein and its molecular weight was about 52kD.The result of Western blot test showed that immunoactivity of the recombinant N protein was the same to that of the natural N protein.
A pair of primers containing BamHI and XhoI sites was designed to amplify n gene of 1149bp from the positive pMD18-TN plasmid of transmissible gastroenteritis virus n gene by PCR. PCR product was subcloned into multiple cloning sites of pET-30a containing 6 His＂ Tag. Recombinant plasmid pET-30a of n gene named pET30a-N was transfected into E. coli BL21 and the bacteria was induced with IPTG at 37℃ . It was demonstrated by SDS-PAGE that recombinant N protein expressed in E. coli BL21 was soluble protein and its molecular weight was about 52kD. The result of Western blot test showed that immunoactivity of the recombinant N protein was the same to that of the natural N protein.
出处
《病毒学报》
CAS
CSCD
北大核心
2006年第2期147-149,共3页
Chinese Journal of Virology
关键词
猪传染性胃肠炎病毒
N基因
原核表达
免疫活性分析
transmissible gastroenteritis virus（TGEV）
n gene
prokaryotic expression
immunoactivity
