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目的 获取具备免疫反应原性的庚型肝炎病毒ＮＳ３ 抗原。方法 从重组质粒Ｉｗｑ２利用ＰＣＲ 反应扩增出ＧＢＶＣ／ＨＧＶ ＮＳ３ 基因片段，克隆至表达载体ｐＰＲＯＥＸ ＨＴａ 后进行核苷酸序列测定，待鉴定正确后经ＩＰＴＧ 诱导重组工程菌，用ＳＤＳＰＡＧＥ 和Ｗｅｓｔｅｒｎ ｂｌｏｔ 对重组蛋白进行鉴定。结果 酶切鉴定和核苷酸序列测定结果表明成功地构建了重组工程菌ｐＨＴＮＳ３／ ＤＨ５α且克隆的基因片段为ＧＢＶＣ／ＨＧＶ ＮＳ３ 基因片段。ＳＤＳＰＡＧＥ 分析诱导表达产物发现在相对分子质量约４３ ８１０处有一条明显的蛋白表达带，占菌体总量的１７ ．７ ％ 。用ＮｉＮＴＡ 亲和层析柱快捷地获得了纯化的重组蛋白。Ｗｅｓｔｅｒｎ ｂｌｏｔ 鉴定发现重组蛋白可与ＧＢＶＣ／ＨＧＶ ＲＮＡ 阳性病人血清发生抗原抗体反应。结论 获得了具备免疫反应原性的ＧＢＶＣ／ＨＧＶ ＮＳ３ 抗原，该重组蛋白可用于ＧＢＶＣ／ＨＧＶ 感染的检测。
Objective To obtain immunogenicity of hepatitis G virus NS3 antigen. Methods The fragment of GBVC / HGV NS3 gene was amplified from the recombinant plasmid Iwq2 by PCR. The nucleotide sequence was determined after cloned into expression vector pPROEX HTa. After identified by IPTG, the recombinant engineering bacteria were induced by SDSPAGE and Western blot analysis of recombinant proteins. Results The results of restriction endonuclease digestion and nucleotide sequencing showed that the constructed recombinant strain pHTNS3 / DH5α was successfully constructed and the cloned gene fragment was GBVC / HGV NS3 gene fragment. SDS-PAGE analysis of the induced expression product found in the relative molecular mass of about 43 810 has a significant protein expression zone, accounting for 17 total bacterial cells. 7%. Purified recombinant protein was obtained rapidly with Ni-NTA affinity chromatography column. Western blot identification found that the recombinant protein with GBV C / HGV RNA positive serum antigen-antibody reaction. Conclusion Obtained with the immunogenicity of GBV C / HGV NS3 antigen, the recombinant protein can be used for GBV C / HGV infection detection.