本文選題：丙型肝炎病毒 + b亞型��； 參考：《病毒學(xué)報》2017年01期

【摘要】：從中國丙型肝炎病毒(HCV)3b亞型感染者克隆分析包膜蛋白編碼基因,并用于HCV 3b亞型假病毒制備。本研究從中國HCV感染血清中篩選克隆到2個3b亞型包膜蛋白編碼基因,序列分析后構(gòu)建表達(dá)質(zhì)粒,以1b(Con1)作為對照,體外轉(zhuǎn)染293T細(xì)胞后分析不同包膜蛋白表達(dá)水平,并制備了HCV 3b亞型假病毒(HCVpp),比較不同HCVpp感染效率。結(jié)果表明:2個3b亞型包膜蛋白編碼基因(C27與C30)與國際3b參考株TrKj的進化關(guān)系較近。C27與C30核苷酸與氨基酸同源性較高(分別為98.5%與98.2%),包膜蛋白編碼區(qū)存在10個氨基酸位點差異,且C27包膜蛋白體外表達(dá)水平明顯高于C30,其中C27可制備出高感染效率的HCV 3b型假病毒,其感染效率明顯高于C30與HCV 1型(Con1)制備的HCVpp。本研究分析了2個HCV 3b亞型感染者包膜蛋白編碼基因序列及表達(dá)特性,并首次獲得了高感染效率的HCV 3b亞型假病毒,為HCV研究及疫苗與藥物研發(fā)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.
【作者單位】： 內(nèi)蒙古醫(yī)科大學(xué)微生物教研室;中國疾病預(yù)防控制中心病毒病預(yù)防控制所;
【基金】：國家自然科學(xué)基金(項目號:81373229) 國家863項目(項目號:2014AA021406) 內(nèi)蒙古自治區(qū)自然科學(xué)基金資助(項目號:2016MS0819)
【分類號】：R373.21
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本文編號：2028432