【摘要】 目的 为研究我国云南1型人类免疫缺陷病毒(Human immunodeficiency virus type 1，HIV-1)流行株外膜蛋白(gp120)的有关抗原表位。方法 采用套式聚合酶链式反应，以来自云南流行区HIV-1感染者的外周血单核巨噬细胞基因组为模板，进行云南流行株外膜蛋白基因(env)片段的扩增，并将env基因片段与原核表达载体pBV220进行重组，构建成质粒pYNenv并在大肠杆菌(E.coli DH10b)中获得表达。采用限制性内切酶分析进行重组质粒的鉴定。结果 含重组质粒的宿主菌经30℃ 20小时培养后，转入42℃培养5小时，经SDS-PAGE蛋白电泳分析有重组蛋白的表达。经Western blot反应证实，该重组蛋白可与来自该流行区的HIV-1感染者血清(含多克隆抗体)发生特异反应。结论 该重组膜蛋白可作为抗原用于HIV-1膜蛋白抗体的检测，并为进一步研究HIV-1 gp120的病理机制奠定了基础。

Abstract The recombinant envelope protein of human immnodificiency virus type 1(HIV-1) of Chinese Yunan epidemic strain was expressed in E.coli for confirming the antigenic domains of the protein. The recombinant prokaryotic expressive plasmid pYNenv was constructed by inserting the env DNA fragment which encode the gp120 of HIV-1 Yunnan strain into pBV220. The env DNA fragment was obtained by nested polymerase chain reaction with the HIV-1 infected patients peripheral blood macrophage and monocyte genomes from Yunan HIV-1 epidemic area as template. The recombinant plasmid pYNenv was confirmed by restriction enzyme analysis. The recombinant protein was expressed in E.coli DH10b by pYNenv after vibration culture at 30℃ for 20 hours then at 42℃ for 5 hours and comfirmed by SDS-PAGE electrophoresis. The protein can specifically react with the serum of HIV-1 infected patient from Yunan epidemic area by Western blot assay. The result showed that the recombinant protein can be used as antigen for detection of HIV-1 membrane glycoprotein antibody as well as for the further study of the role of envelope protein in pathology.

     ......