J亚群禽白血病病毒gp85基因的克隆与表达

Cloning and Expression of gp85 Gene of Avian leukosis virus Subgroup J

构建J亚群禽白血病病毒gp85基因的原核表达载体,并在大肠埃希菌中获得表达。根据原核表达载体pET30的酶切图谱设计引物,以重组质粒pBS-T-env为模板,扩增gp85基因。将此段基因克隆到表达载体pET30上,转化大肠埃希菌BL21,用IPTG诱导表达,经SDS-PAGE分析,表达出约为70ku的融合蛋白。获得了gp85基因的表达产物,为研制ALV-J的特异性抗血清及国内ALV-J病毒分子诊断试剂盒奠定了基础。

To construct the prokaryotic expression vector for gp85 gene of avian leukosis virus subgroup J and express the fusion protein in E. coli. According to the prokaryotic expression vector restriction map pET30, a pair of primers were designed to amplify the gp85 gene and the recombinant plasmid pBS-T-env was used as templates for PCR reaction. The gp85 gene was cloned into the expression vector pET30, transformed in E. coli BL21 and induced by IPTG. SDS-PAGE analysis showed that the fusion protein was about 70 ku in size,indicatin that Obtained the expressive product of the gp85 gene was obtained, it lays a basis for developing the special antiserum of ALV-J and the domestic ALV-J molecular diagnostic reagent.