期刊论文

中文

中国预防兽医学报

1008-0589

2010

32

2

126-130

基金项目：湖南省科技重大专项（2007FJ1003） 湖南省农业大学***基金（04YJl03）

本校为第一机构

为表达猪巨细胞病毒(PCMV)gB基因,本研究根据GenBank登录的PCMV gB基因序列设计1对引物,以感染PCMV猪肺细胞总DNA为模板,采用PCR扩增得到gB基因片段,克隆于pMD-18T并进行核苷酸序列测定.利用DNAStar分析gB蛋白的抗原表位,选择抗原表位富集的2个基因片段(命名为PCMVgB1和PCMVgB2)分别克隆到原核表达载体pET-32a(+)中,构建重组表达质粒并转化Rosetta(DE3)宿主菌.结果显示:扩增的gB基因与GenBank登录的PCMV gB基因的核苷酸同源性在97.6%～98.9%之间,推导的氨基酸同源性在97%～98.6%之间.经IPTG诱导的含pET-gB1和pET-gB2重组质粒的宿主菌可表达重组gB1和gB2蛋白,SDS-PAGE显示分子量约为62ku和36 ku....

The glycoprotein B （gB） gene encoding structural protein of porcine cytomegalovirus （PCMV） was amplified by PCR using primers derived from the published PCMV gB gene sequence, cloned and sequenced. The gB gene shared 97.6 %-98.9 % sequence identity at nucleotide level and 97 %-98.6 % identity at amino acid level with PCMV gB reference genes in GenBank. Two immunodominant regions of PCMV gB1 and PCMV gB2 were identified by DNAStar and were subcloned into prokaryotic expression vector pET-32a（＋）. The resulting plasmids of pET-gB1 and pET-g...