中国株HGV NS3 区基因的原核表达及其产物的抗原性分析

Expression of nonstructural region 3 T5HZ gene of the Chinese HGV and analysis of the antigenicity of the recombinant proteinsX

目的研究ＨＧＶＮＳ３区基因产物的抗原性，并探讨ＮＳ３蛋白在血清学检测中的应用。方法将中国株ＨＧＶＮＥ３区的３个基因片段分别克隆到ｐＲＳＥＴＢ和ｐＲＳＥＴＣ质粒载体中，构建成原核表达载体。ＩＰＴＧ诱导下，在大肠杆菌ＢＬ２１中高效表达，获得３个重组蛋白，用Ｗｅｓｔｅｒｎｂｌｏｔ和ＥＬＩＳＡ法分别对表达产物进行分析。结果所构建的表达载体均得到高效表达，得到的重组蛋白ＰＡ、Ｐ３和Ｐ４的分子量分别为４２０００，３００００和２４０００，在Ｗｅｓｔｅｒｎｂｌｏｔ和ＥＬＩＳＡ反应中均可被ＨＧＶ阳性血清识别，其中ＨＧＶＮＳ３区Ｎ端的基因产物抗原性较强。结论中国株ＨＧＶＮＳ３区的Ｎ端存在优势的抗原决定簇，其基因产物有较好的抗原性。

Objective This study is to analyze the antigenicity of the NS3 proteins of Chinese HGV and their potential use in the serological diagnosis. Methods All three gene fragments of NS3 region of Chinese HGV were cloned into the pRSET vectors to construct recombinant plasmids. In E.coli BL21, all three recombinant plasmids achieved a high expression level with induction of IPTG. The expressed products were analyzed with Western blot and ELISA. Results The recombinant protein PA, P3 and P4 have a molecular weight of 42 000, 30 000 and 24 000, respectively. They all could react with HGV positive sera in Western blot and ELISA. Among them, the protein that covers the N terminal of NS3 region of HGV had a stronger reaction with HGV positive sera than the other two proteinsdid. Conclusion The N terminal in the NS3 region of Chinese HGV includes an dominant antigenic determinant, and its gene product has relatively strong antigenicity.