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在大肠杆菌中， 利用ｐＤＯＧ 载体来源的表达载体ｐＤＧａｇＥｎｖ ， 高效表达了相对分子质量（ Ｍｒ） 为４６ ５００（ｐ４６） 的重组ＨＩＶ１Ｇａｇ／ Ｅｎｖ 融合抗原（Ｇａｇ２０９ ～４３７ ＋ Ｅｎｖ４７４ ～５１８ ＋ Ｅｎｖ５４８ ～６７５） 。表达产物同时还包括：Ｍｒ ～２８ ５００ ， Ｍｒ ～２２ ０００ 与 Ｍｒ ～１９ ０００ ３ 种（ｐ２８ ，ｐ２２ 和ｐ１９） 蛋白。此４ 种表达产物均存在于包涵体中，不能被８ ｍ ｏｌ／Ｌ 尿素溶解。Ｗｅｓｔｅｒｎ 印迹分析表明，４ 种重组蛋白均能与ＨＩＶ１ 阳性血清发生特异反应；其中ｐ４６ 与ｐ２８ 蛋白能与抗ＨＩＶ１ｐ２４ 反应，而ｐ２２ 与ｐ１９ 蛋白则不与其起反应。其中ｐ１９ 蛋白对应于Ｅｎｖ４８２ ～５１８ ＋ Ｅｎｖ５４５ ～６７５ ，ｐ２２ 蛋白可能是从Ｇａｇ／ Ｅｎｖ 表达质粒的ｇａｇ 基因内部的甲硫氨酸（ＡＴＧ４２３） 起始的翻译产物。将包涵体充分洗涤后，利用制备电泳方法纯化了ｐ４６ ，ｐ２２／ｐ１９ 及ｐ１９ 蛋白。将后两种纯化蛋白质以２ ｍ ｇ／Ｌ 的浓度包被ＥＬＩＳＡ 板，检测了国家卫生部药品生物制品检定所提供的ＨＩＶ１／ ＨＩＶ２ 质检血清。结果可检出其中包含的所有１８ 份ＨＩＶ?
In E. coli, the recombinant HIV-1 Gag / Env fusion antigen (Gag209-437 + Env474-518 + Env548-675) with the relative molecular mass of 46 500 (p46) was efficiently expressed using the expression vector pDGagEnv of pDOG vector ). The expression product also includes 3 proteins of Mr ~ 28 500, Mr ~ 22 000 and Mr ~ 19 000 (p28, p22 and p19). The four expressed products are present in the inclusion body, can not be 8mol / L urea dissolved. Western blot analysis showed that all four recombinant proteins reacted specifically with HIV-1 positive serum. Among them, p46 and p28 protein reacted with anti-HIV-1p24, while p22 and p19 protein did not react with it. The p19 protein corresponds to Env482-518 + Env545-675, and the p22 protein may be the translation product starting from methionine (ATG423) inside the gag gene of Gag / Env expression plasmid. After the inclusion bodies were sufficiently washed, p46, p22 / p19 and p19 proteins were purified by preparative electrophoresis. The latter two purified proteins were coated with ELISA plate at a concentration of 2 m g / L and tested for HIV-1 / HIV-2 quality sera provided by the National Institute of Biological Control of Medicines. As a result, all 18 HIVs contained in it can be detected?