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丙型肝炎病毒(HCV)是一正链 RNA 病毒,基因组全长约9400bp,由结构区(C 区、E 区)、非结构区(NS_1～NS_5)及3′端和5′端非编码区组成。非结构5区(NS_5)靠近3′末端,长约3100bp,依5′至3′的顺序分为 NS_(5a)及 NS_(5b)区。据报道 NS_(5a)在基因分型中有重要作用。NS_(5a)抗体的检测对改善诊断试剂灵敏度有重要作用。NS_(5b)区域则主要是依赖于 RNA 的 RNA 聚合酶编码区域。目前,国内已有关于 NS_(5a)基因表达的报道,但关于 NS_(5b)基因区域的表达还没有正式文献。我们用含多个连续组氨酸的表达质粒,在大肠杆菌中表达了 NS_(5b)基因区段。一、资料与方法1.质粒、菌株及试剂:宿主菌E.coli 06由法国巴斯德研究所惠赠。表达质粒 pTric.His 由我所沈月雷硕士提供。含 NS_5及
Hepatitis C virus (HCV) is a positive-stranded RNA virus with a genome of about 9,400bp in length and consists of the structural regions (C and E), the non-structural regions (NS_1 and NS_5) and the 3 ’and 5’ noncoding regions composition. The non-structural 5 region (NS_5) is near the 3 ’end, about 3100 bp in length and is divided into NS_ (5a) and NS_ (5b) regions in the order of 5’ to 3 ’. It has been reported that NS_ (5a) plays an important role in genotyping. The detection of NS5a antibody plays an important role in improving the sensitivity of diagnostic reagents. The NS_ (5b) region is mainly dependent on RNA RNA polymerase coding region. At present, NS5a gene expression has been reported in China, but there is no official literature on the expression of NS5b gene region. We expressed the NS_ (5b) gene segment in E. coli using an expression plasmid containing multiple consecutive histidines. I. Materials and Methods 1. Plasmids, Strains and Reagents: The host strain E. coli 06 was kindly donated by the Institut Pasteur. The expression plasmid pTric.His was provided by my master, Shen Lei. With NS_5 and