【摘要】 目的提高人免疫缺陷病毒Ⅰ型（ＨＩＶ－１）包膜糖蛋白ｇｐ１２０基因在原核系统中的表达量。方法采用聚合酶链反应（ＰＣＲ）技术扩增出５６０ｂｐ的ＨＩＶ－１ＬＡＶ株ｇｐ１２０Ｎ端基因片段，经ＥｃｏＲⅠ及ＳａｌⅠ酶切后插入高效表达载体ｐＥＴ２８ａ，得到重组质粒ｐＥＴ１２０，并转化表达宿主菌ＢＬ２１（ＤＥ３），经诱导高效表达出ＨＩＶ－１ｇｐ１２０基因片段。结果间接酶联免疫吸附试验（ＥＬＩＳＡ）及Ｗｅｓｔｅｒｎｂｌｏｔ实验表明，表达产物具有良好的抗原性及特异性。ＳＤＳ－ＰＡＧＥ电泳分析结果表明，ｇｐ１２０表达量占总菌体蛋白的５０％。结论在原核系统中高效表达了ＨＩＶ－１ｇｐ１２０基因更多还原

【Abstract】 In order to improve protein expression and to produce good and cheap diagnostic antigen,the gene fragment (560bp)in N-terminal of gp120 of HIV-1 LAV strain was amplified by PCR.After digested by EcoRⅠ and SalⅠ,the fragment was cloned into a high-level expression vector pET28a.The recombinant plasmid pET120 transfecting BL21(DE3)produced the protein with high-level expression in the host cell BL21(DE3),which was further proved having good antigenicity and high specificity by indirect ELISA and Western-blot assay.The proteiu expressed was about 50% of the total bacterial protein by SDS-PAGE electrophoresis test.It was highly expressed in the prokaryotic expression system.更多还原