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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。
The gag gene and env gene of subtype B of HIV-1 were inserted into prokaryotic expression vector p ET2 8a and p ET2 8c to obtain recombinant expression plasmids p ETG, p ETM1, p ETM2 and p ETM3 . P ETG was transformed into host strain BL21 (DE3) plys S, induced by IPTG, and analyzed by SDS-PAGE. The protein expression increased gradually with the induction time, reaching the maximum 4 h. The results of gel scanning showed that the protein expression accounted for 10.3% of the total bacterial protein. Western blotting showed that the recombinant protein reacted specifically with the HIV-1 B subtype sera from Chinese endemic strains. The recombinant expression plasmids p ETM1, p ETM2 and p ETM3 showed specific bands only in the Western blot with gp1 20 mAb, but no band was detected by SDS-PAGE.