含中国流行株HIV-1Gag-gp120融合蛋白的重组鸡痘病毒的构建  

Construction of recombinant fowlpox virus with HIV-1Gag-gp120 fusion protein

作　　者：金宁一[1] 罗坤[2] 方厚华[1] 王莉馨[1] 郭志儒[1] 郭焱[1] 秦云龙[1] 安汝国[3] 殷震[1] 

机构地区：[1]解放军军需大学基因工程实验室,长春130062 [2]浙江大学免疫学研究所,杭州310006 [3]白求恩医科大学地方病研究所,长春130021

出　　处：《中国免疫学杂志》2001年第8期408-410,共3页Chinese Journal of Immunology

基　　金：国家杰出青年基金资助 (基金批准号 3982 5 1197)

摘　　要：目的 :用重组鸡痘病毒表达中国流行株HIV 1Gag gp12 0融合蛋白。方法 :在以鸡痘病毒 2 82E4株为基础构建的重组表达质粒pUTAL的ATI P7 5复合启动子下游 ,插入编码中国流行株HIV 1Gag gp12 0嵌合基因 ,构建了重组表达质粒pUTALGP。重组质粒与FPV共转染CEF细胞 ,进行了同源重组。通过PUdR加压筛选 ,X gal染色 ,获得重组鸡痘病毒vUTAL GP。结果 :经Westernblot检测表明 ,重组病毒表达了Gap gp12 0融合蛋白 ,其分子量分别为 110× 10 3、6 9× 10 3、48× 10 3、2 4×10 3。电镜观察可见Gag蛋白在CEF细胞中形成的反转录病毒样粒子。重组病毒免疫小鼠 ,能够诱导HIV 1特异的血清抗体反应。结论 :重组鸡痘病毒成功的表达了Gag gp12 0融合蛋白 ,并进行了正确的加工。Objective:To express Gag gp120 fusion protein in the recombinant fowlpox virus.Methods:The Gag gp120 gene of HIV 1 was inserted downstream of the combined promoter in pUTAL,an expressing plasmid was constructed.By transfecting the plasmid into CEF cells pre infected with FPV 282E 4 strain,a recombinant fowlpox virus vUTALG was obtained by selecting in the presence of BUdR and subsequent blue plaque screening.Results:Western blot showed that the recombinant virus expressed the Gag gp120 fusion protein in the infected CEF cells.The virus like particles formed by Gag protein were observed under electron microscope.Mice were immunized with the recombinant virus.The results showed the recombinant virus could induce HIV 1 specific antibody response.Conclusion:The recombinant fowlpox virus could express the Gag gp120 fusion protein.

关 键 词：HIV-1 Gag-gp120 重组鸡痘病毒 

分 类 号：R373.9[医药卫生—病原生物学]