猪繁殖与呼吸综合征病毒Hn-1/06株GP2蛋白的原核表达  

Prokaryotic Expression of GP2 Protein of Porcine Reproductive and Respiratory Syndrome Virus Hn-1/06 Strain

作　　者：刘明莉[1] 李伟娟[1] 王中明[1] 邱璜[1] 卢高峰[1] 夏平安[1] 李素平[1] 崔保安[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《江西农业学报》2009年第3期4-7,共4页Acta Agriculturae Jiangxi

摘　　要：根据GenBank中PRRSV美洲株ATCC VR2332的ORF2基因序列,利用Primer5.0软件设计合成了一对特异性引物,经RT-PCR从河南分离株Hn-1/06株扩增得到了大小为771 bp的片段,并将扩增的片段插入pTG19-T载体进行测序而获得含有ORF2的阳性克隆pTG19-T-GP2。将此基因亚克隆到原核表达载体pGEX-6p-1,经筛选获得了阳性重组质粒pGEX-GP2,进而在IPTG的诱导下成功表达,获得了大小约为45 kD的融合蛋白,纯化后经Western blot检测证实表达的融合蛋白具有良好的生物学活性。A pair of specific primers of porcine reproductive and respiratory syndrome virus （PRRSV） ORF2 were designed and synthesized according to the published gene sequence of PRRSV from GenBank （Accession number： PRU87392）, and ORF2 gene of PRRSV Hn - 1/06 strain was obtained by RT - PCR method. The RT - PCR product was directly cloned into pTG19 - T vector and this recombinant plasmid was identified by enzyme digestion and DNA sequencing. The positive clone was designated pTG19 - T - GP2. After being cloned into the prokaryotic expression vector pGEX - 6p - 1, the ORb2 gene was successfully obtained with the induction of IPTG. Western - blot test result confirmed that the expressed fusion protein could specifically react with antiserum against PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 GP2蛋白 原核表达 

分 类 号：S828[农业科学—畜牧学]