羊痘病毒P32基因与羊CD58基因共表达载体的构建

摘要

为了开发出一种可行的羊痘基因疫苗,尝试用羊P32基因与CD58基因建立共表达载体.试验根据Gen-Bank中收录的羊痘病毒(GPV)P32和羊CD58基因组序列,设计了扩增 GPV P32基因和CD58基因的特异性引物,运用PCR技术从 GPV中扩增出P32基因,从羊的外周血中扩增出CD58基因,并将其分别克隆到 pMD18-T载体,测序正确.将P32基因去掉后端疏水区一段与CD58基因先后克隆至载体 pBudCE4.1,并转化大肠埃希菌JM109,提取质粒通过酶切鉴定和测序正确,且读码框正确.说明成功获得了真核共表达质粒 pBudCE4.1/CD58/P32.%In order to develop a gene capripox vaccine,this study attempts to establish a co-expression vector contain the goatpox virus P32 gene and sheep CD58 gene.Based on the sequences of goatpox virus (GPV)P32 and the sequences of sheep CD58 gene published in the GenBank,designed to amplify GPV P32 and CD58 gene-specific primers,the P32 gene was amplified by PCR from GPV and amplification of CD58 gene from peripheral blood of sheep,cloning its to pMD18-T carrier,sequencing is correct.The P32 gene removed some backend hydrophobic region and CD58 gene has cloned into the vector pBudCE4.1,and transformed into E.coli JM109,plasmid by restriction enzyme digestion,sequencing and proper reading frame correct,successful eukaryotic co-expression plasmid pBudCE4.1/CD58/P32.