鸡传染性贫血病毒VP2基因的克隆和原核表达

Cloning and prokaryotic expression of chicken anemia virus VP2 gene

从组织病料中提取到的鸡贫血病毒核酸经PCR扩增得到vp2基因,并将其克隆到表达载体pET32a上,重组菌经IPTG诱导,SDS-PAGE分析,结果VP2基因在大肠杆菌中成功表达。以表达产物免疫小鼠4次,制备了抗CAVVP2的多克隆抗体。通过对CAV感染的MSB1细胞做间接免疫荧光试验(IFA),结果为阳性。这表明表达产物保留了CAV相关的抗原性。为进一步研究CIA临床诊断奠定了基础。

Chicken anemia virus genome extracted from liver tissue, was amplified with PCR and VP2 gene was gained, then inserted into pET32a vector and transformed into competent E. coli cell plys. The recombinant bacteria was induced by IPTG and analyzed with SDS-PAGE. The result showed that gene encoding VP2 was expressed in E. coli at high level. The anti-CAV polyclonal antibody was produced in mice immunized four times with the expressed protein ,and it showed positive reaction with CAV-infected MSB 1 cells in indirect immunofluorescent assay （IFA）. This result showed that the expressed protein retained the CAV-related antigenicity can will be used to the development of clinical diagnosis of CIA.