丙型肝炎病毒JFH1适应性突变株全长cDNA的构建

Construction of Full-length Complementary DNAs of Hepatitis C Virus JFH1 Strains with Adaptive Mutations

目的构建丙型肝炎病毒JFH1全长c DNA克隆,获取突变位点信息。方法应用长片段RT长片段RT-PCR技术对不同时期传代的JFH1病毒株进行分段扩增。通过共同酶切位点连接成9.7Kb从DNA片段,与p Blue Script2ks(+)形成克隆载体,进一步测序分析。结果获取丙型肝炎病毒JFH1系列适应性突变株全长c DNA模板,共发现23个单核苷酸突变。结论丙型肝炎病毒JFH1系列存在单核苷酸突变位点,为进一步了解HCV JFH1适应性突变位点功能奠定基础。

Objective To construct the full length complementary DNAs of HCV JFH1 strains with adaptive muta- tions. Methods Three HCV gene fragments （ 1.5 kb,4.1 Kb and 4.1 kb） were amplified by long RT-PCR, then fused and connected together to produce a 9.6 kb complete HCV genome,which was further cloned into a pBlueScript2ks（ ＋ ） vector. The cDNAs were analyzed by sequencing and compared with the original HCV JFH1 strain. Results Successful got the adaptive HCV JFHI＇s full length cDNAs and found 23 single-nucleotide mutations. Conclusion Our findings will provide further insights into the function of adaptive mutations of the HCV JFHI.