Ⅰ型人免疫缺陷病毒(HIV-1)中国流行株B亚型Gag和Env蛋白在大肠杆菌中的表达

Expression of Gag Protein and Env Protein of HIV-1 Subtype B in E.coli

将中国流行株 型人免疫缺陷病毒 ( HIV-1 ) B亚型 gag基因和 env基因定向插入原核表达载体p ET2 8a和 p ET2 8c,获得重组表达质粒 p ETG、p ETM1、p ETM2和 p ETM3。将 p ETG转化表达宿主菌 BL2 1( DE3 ) plys S,用 IPTG诱导 ,SDS-PAGE电泳分析有重组蛋白表达。该蛋白表达量随诱导时间的延长而逐渐增加 ,4 h达到最大值。凝胶扫描结果显示 ,该蛋白表达量占菌体总蛋白的 1 0 .3 %。Western blotting检测 ,该重组蛋白可与中国流行株 HIV-1 B亚型阳性血清发生特异反应。重组表达质粒 p ETM1、p ETM2和 p ETM3仅在蛋白印迹中与 gp1 2 0单抗发生特异性反应而出现特异显色带 ,SDS-PAGE电泳分析未见表达蛋白带。

In order to study the molecular biology of HIV 1 and to develop diagnostic reagent for AIDS in China. Four recombinant expression plsmids pETG,pETM1,pETM2 and pETM3 were constructed by inserting the gag gene and env gp120 fragments of HIV 1 subtype B into the vectors pET28a and pET28c. Then the expression plasmids were transformed into expressing host cell BL21(DE3)plysS ,induced by IPTG.The results of SDS PAGE and Western blotting assay demonstrated that the protein with a molecular of 55 000 expressed by the recombinant plasmid pETG in E.coli can be specifically recognized by antiserum from the infected individuals of HIV 1 subtype B and densitometric scan analysis showed the contents of expressed HIV 1 gag protein was 10.3% of the total amount of the host cell protein.The recombinant expression plasmids pETM1,pETM2 and pETM3 can specifically response to monoantibody of gp120 indicting may be used as diagnostic reagents for HIV infection in China.