中国HCV3b型包膜蛋白编码基因的克隆分析与假病毒制备

发布时间：2018-06-16 23:12

  本文选题：丙型肝炎病毒 + b亚型　； 参考：《病毒学报》2017年01期

【摘要】：从中国丙型肝炎病毒(HCV)3b亚型感染者克隆分析包膜蛋白编码基因,并用于HCV 3b亚型假病毒制备。本研究从中国HCV感染血清中筛选克隆到2个3b亚型包膜蛋白编码基因,序列分析后构建表达质粒,以1b(Con1)作为对照,体外转染293T细胞后分析不同包膜蛋白表达水平,并制备了HCV 3b亚型假病毒(HCVpp),比较不同HCVpp感染效率。结果表明:2个3b亚型包膜蛋白编码基因(C27与C30)与国际3b参考株TrKj的进化关系较近。C27与C30核苷酸与氨基酸同源性较高(分别为98.5%与98.2%),包膜蛋白编码区存在10个氨基酸位点差异,且C27包膜蛋白体外表达水平明显高于C30,其中C27可制备出高感染效率的HCV 3b型假病毒,其感染效率明显高于C30与HCV 1型(Con1)制备的HCVpp。本研究分析了2个HCV 3b亚型感染者包膜蛋白编码基因序列及表达特性,并首次获得了高感染效率的HCV 3b亚型假病毒,为HCV研究及疫苗与药物研发提供了有效工具。
[Abstract]:The envelope protein encoding gene was cloned from Chinese hepatitis C virus (HCV) subtype 3b infection and used for preparation of HCV 3b pseudovirus. In this study, two 3b subtype envelope protein coding genes were screened and cloned from Chinese HCV infected serum. After sequence analysis, expression plasmids were constructed, and the expression levels of different envelope proteins were analyzed after transfection of 293T cells in vitro. HCV 3b subtype pseudovirus was prepared and compared with different HCV pp infection efficiency. The results showed that the evolutionary relationship between two 3b subtype envelope protein coding genes (C27 and C30) and the international 3b reference strain TrKj was closer. C27 and C30 had higher homology of nucleotide and amino acid (98.5% and 98.2%, respectively). There were 10 amino acid loci difference in the coding region of the envelope protein. The expression level of C27 envelope protein in vitro was significantly higher than that of C30. C27 could produce HCV 3b pseudovirus with high infection efficiency, and its infection efficiency was significantly higher than that of HCV-pp1 prepared by C30 and HCV-1. In this study, the sequence and expression characteristics of two envelope protein coding genes in HCV 3b subtype infected patients were analyzed, and the highly efficient pseudoviruses of HCV 3b subtype were obtained for the first time, which provided an effective tool for HCV research, vaccine and drug development.
【作者单位】： 内蒙古医科大学微生物教研室;中国疾病预防控制中心病毒病预防控制所;
【基金】：国家自然科学基金(项目号:81373229) 国家863项目(项目号:2014AA021406) 内蒙古自治区自然科学基金资助(项目号:2016MS0819)
【分类号】：R373.21
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本文编号：2028432