【摘要】 根据绵羊肺腺瘤病毒(SJRV) NM株基因组全序列,设计了 1 对包含囊膜(env)基因在内的引物,并在5′端分别设计了HindⅢ和 BamHⅠ酶切位点,利用 PCR技术扩增 env基因,通过连接反应将其克隆于pET 30b表达载体上。序列分析表明,env基因全长 1 836 bp,编码 612 个氨基酸残基;构建表达重组质粒env pET 30b,转化大肠埃希氏菌 BL21,经 IPTG于 37 ℃诱导培养,SDS PAGE电泳分析表明,表达的env基因融合蛋白分子质量约为69 ku。更多还原

【Abstract】 The envgene was amplified with a pair of primers with HindⅢ and BamHⅠsites on the 5′-terminal, which was designed according to the sheep Jaagsiekte retrovirus NM strain(SJRV-NM) genome sequence, by PCR and then cloned in the vector pET-30b. The sequence analysis indicated that the env gene encoding 612 amino acid residues was consisted of 1836bp nucleotides. The expression recombiant plasmid was constructed in vitroby inserting the env gene into the vector pET-30b, and then transformed into E.coli BL21.The bacteria was induced by IPTG. The SDS-PAGE analysis showed that the env fusion protein had been expressed and its molecule is 69ku.更多还原