HCV核心区基因的截短及其表达 (pdf)

    【摘要】 目的 高效稳定地表达HCV的核心抗原。方法 设计一对特异性引物，以含HCV全长基因的质粒pBR TM/HCV1-3011为模板，扩增出HCV截短的核心抗原(1-120aa)的基因，克隆于表达载体pGEX-4T3,并转化于大肠杆菌BL21,IPTG诱导HCV截短的核心抗原的表达，表达产物经过SDS-PAGE,Western-blot,ELISA等一系列鉴定试验进行了分析。结果 成功地构建了HCV核心抗原高效稳定表达的重组质粒，表达产物具有高度特异性和良好的抗原活性。结论 该抗原的获得为研制诊断用HCV抗原奠定了基础。

    【关键词】 HCV；核心区基因；截短；表达

    Truncation and expression of HCV Core gene

    PENG Xiang-bing,SANG Ai-jun,HUANG Shi-he,et al.Wuhan Institute of Biological Products,Wuhan 430060,China

    【Abstract】 Objective To express HCV Core antigen hightly and stably.Methods A pairs of specific PCR primers were designed and synthesized.Truncated HCV Core gene was amplified from the plasmid pBR TM/HCV1-3011 by PCR.The gene fragement was cloned into pGEX-4T3,then transformed to E.coli BL21and expressed under induction of IPTG. The expressed product was analyzed by SDS-PAGE,Western-blot and indirect ELISA,respectively.Results A recombinant plasmid pGEX-TP1.1for stable and hight expression was constructed successfully. The expressed product showed high specificity and good antigenicity.Conclusion The expression of truncated HCV Core antigens laid a foundation for development of HCV diagnostis use antigens. ......