【摘要】 目的表达人免疫缺陷病毒1型C亚型核心蛋白Gag,为HIV感染的诊断和疫苗的研制奠定基础。方法通过PCR扩增获得HIV1gag基因片段,并将其克隆到原核表达载体pGEX5X1的tac启动子下游,构建重组表达质粒,转化大肠杆菌BL21(DE3),IPTG诱导表达,以SDSPAGE和Westernblot分析验证其表达产物。结果表达产物是相对分子质量为82000的GST融合蛋白,且能与抗p24单克隆抗体发生特异性反应。结论重组Gag蛋白在大肠杆菌中得到了有效表达,且具有良好的抗原性,可进一步用于HIV感染及动物免疫等方面的研究。更多还原

【Abstract】 Objective To express the core protein (Gag) of HIV-1 subtype C and lay a foundation of diagnosis of HIV infection and development of HIV vaccine.Methods The Gag gene fragment of HIV-1 was amplified by PCR,inserted downstream to the tac promoter of prokaryotic expression vector pGEX-5X-1.The constructed recombinant plasmid was transformed to E.coli BL21(DE3) for expression under induction of IPTG.The expressed product was identified by SDS-PAGE and Western blot.Results The GST fusion protein,with a relative molecular weight of 82 000,was expressed in E.coli and recognized by McAb to p24.Conclusion Recombinant Gag protein was effectively expressed in E.coli and showed good antigenicity.The expressed protein might be used for the detection of HIV infection and the study on immunization in animals.更多还原