中文摘要：

目的：扩增丙型肝炎病毒（Hepatitis Cvirus,HCV）P7蛋白基因,克隆到多个原核表达系统,观察P7蛋白在体外表达的情况,获取具有生物学活性的HCVP7重组蛋白。方法：设计扩增全长HCVP7基因片段的特异引物,以H/FL质粒DNA（含HCV1acDNA全长序列）为模板,通过PCR扩增全长HCV P7基因,定向克隆到pQE30、pGEX 4T-2、pET32a（＋）3种不同类型的原核表达载体中。将重组后包含HCVP7基因的原核表达载体转化表达型大肠杆菌BL21（DE3）pLysS或SG13009（PREP4）,并做异丙基硫化-β-D-半乳糖苷（Isopropyl-beta-D-thiogalactoside,IPTG）诱导表达,通过SDS-PAGE和Western blot鉴定HCV P7蛋白原核表达的情况。结论：获得可溶性的重组蛋白GST-HCVP7和Trx-HCVP7,为进一步研究P7蛋白的生物学功能奠定基础。

英文摘要：

Objective： To amplify HCV P7 gene and clone it into several prokaryotic expression systems,then investigate the effects of these working systems and harvest HCV P7 protein with biologic activity.Methods： Design Special primers were designed for the amplification of HCV P7 complete DNA sequence and amplify P7 DNA fragments were amplified with H/FL plasmid as template by PCR.The obtained P7 DNA fragments were directly cloned into pQE30,pGEX 4T-2 and pET32a（＋） prokaryotic expression vectors and transform these recombinant plasmids were transformed into E coli.strain BL21（DE3） pLysS or SG13009（PREP4）,and IPTG was added to induce recombinant E coli.expression.The expression products were identified with SDS-PAGE and Western blot.Results： There was no track of non-fusion HCV P7 protein expression from recombinant E coli,including pQE30/HCV P7 or pET32a（＋）-Nde/HCV P7 plasmid.But the study succeeded in the expression and purification of recombinant HCV P7 proteins,Both are all soluble and biologic activity,and are expected to use extensively in anti-HCV virus research.