HIV-2 env基因在大肠杆菌和重组痘苗病毒中的表达

Expression of HIV 2 env Gene in E.coli and Recombinant Vaccinia Virus

将编码人Ⅱ型免疫缺陷病毒（ＨＩＶ－２）ｅｎｖｇｐ１２０（Ｅ１）和ｇｐ３６（Ｅ２）分别克隆到原核高效表达载体ｐＥＴ１７ｂ、ｐＢＶ２２０和真核表达载体ｐ１０、ｐ１６中，构建成８个重组质粒。经ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ分析证明，在原核表达系统成功地表达了ＨＩＶ－２ｅｎｖ融合蛋白，表达的蛋白分子量分别为３５０００、７００００和５００００，而原核表达质粒ｐＥＴＥ１在ＳＤＳ－ＰＡＧＥ凝胶的３５０００处可见明显的额外蛋白带。经薄层扫描测定，原核系统表达的Ｅｎｖ蛋白（ｇｐ１２０）相对含量为５．８％。在真核细胞中表达的Ｅｎｖ蛋白经Ｗｅｓｔｅｒｎｂｌｏｔ检测分析，分子量分别为６００００、５７０００和４２０００。上述表达的Ｅｎｖ蛋白均能与ＨＩＶ－２阳性血清发生特异性反应。

Eight recombinant plasmids were constructed by inserting the HIV 2 env gp120(E1) and gp36 (E2) genes into pET17b, pBV220, p10 and p16 vectors. The results of Western blot assay demonstrated that HIV 2 Env proteins were able to express in E.coli BL21 (DE3) and BHK 21 cells, and could specifically response to HIV 2 positive serum. The weight of procaryotic products were 35 000 , 70 000, 50 000 and eucaryotic products were 60 000, 57 000 and 42 000. SDS PAGE and densitometric analysis showed that the content of pETE1 was 5.8%. It was proved that the recombinant vaccinia viruses were able to induce HIV 2 specific antibody in mice.