中国HCV3b型包膜蛋白编码基因的克隆分析与假病毒制备  

Characterization of Glycoproteins and Preparation of HCV “Pseudoparticles” with High Infectivity of Subtype 3b from Chinese Patients

作　　者：陶格斯[1,2] 叶飞[2] 邓瑶[2] 张玲[2] 卢莎[1] 博晓真[1] 李从力[1] 沈晓玲[1] 谭文杰[2] TAO Gesi YE Fei DENG Yao ZHANG Ling LU Sha BO Xiaozhen LI Congli SHEN Xiaoling TAN Wenjie(Department of Microbiology, Inner Mongolia Medical University, Hohhot 010059, China National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention Key Laboratory of Medical Virology, Ministry of Health, Beijing 102206, China)

机构地区：[1]内蒙古医科大学微生物教研室,呼和浩特010059 [2]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《病毒学报》2017年第1期56-60,共5页Chinese Journal of Virology

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)

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。We wished to characterize the envelope protein coding gene of the hepatitis-C virus （HCV） from Chinese patients and prepare HCV pseudoparticles （HCVpp） of subtype 3b. Two of the HCV genotype 3b envelope protein coding genes （C27, C30） were cloned from Chinese HCV patients followed by sequencing analyses. Then, two envelope protein expression plasmids were constructed and characterized by western blotting. HCVpp were prepared and target cells infected in vitro. Results showed that the sequences of nucleotides and amino acids from two HCV subtype 3b envelope protein encoding genes （C27, C30） had high homology （98. 5% and 98. 2%, respectively） and had a close phylogenetic relationship with the international reference strain 3b TrKj. Ten amino-acid sites were substituted in the envelope protein coding region of C27 and C30. Expression of the C27 envelope protein was significantly higher than that of HCV subtype 1 （Conl） and C30. The corresponding HCVpp infectivity in vitro was also significantly different, whereby C27 could be used to prepare HCVpp subtype 3b with high infectivity. In conclusion, two envelope protein encoding genes of HCV subtype 3b were sequenced and their expression in vitro investigated. This is the first report on preparation of HCVpp subtype 3b with high infectivity. This study might provide an effective tool for HCV research and development of a vaccine for HCV.

关 键 词：丙型肝炎病毒 3b亚型 包膜蛋白 假病毒颗粒 感染效率 

分 类 号：R373.2[医药卫生—病原生物学]