摘要
为获得蓝舌病病毒(BTV)VP7基因的原核表达蛋白,本实验采用RT-PCR方法扩增出了血清1型BTV的VP7基因,并克隆到原核表达载体pMAL-c2X中,构建了重组质粒pMAL-VP7。将重组质粒转化TB1感受态细胞,以0.5mmol/L的IPTG进行诱导表达,SDS-PAGE电泳结果显示融合蛋白MBP-VP7以可溶形式存在,分子质量约为90ku。以纯化的重组蛋白作为包被抗原,初步建立了检测BTV血清抗体的间接ELISA诊断方法,为今后进一步开展BTV诊断研究奠定了基础。
In this study, the VP7 gene of serotype 1 bluetongue virus was amplified by RT-PCR and cloned into prokaryotic expression vector pMAL-c2X. The recombinant plasmid was transformed into E. coli TB1 cells and fusion protein （MBP-VPT） produced after induced with 0.5 mmol/L IPTG. The expressed protein was about 90 ku and existed in soluble form. Indirect ELISA was established using purified MBP-VP7 protein as antigen.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2009年第12期925-928,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家863项目(No.2007AA02Z406)
