赵 方,刘映霞,李 刚.丙型肝炎病毒JFH1适应性突变株全长cDNA的构建.[J].中南医学科学杂志.,2015,43(3):253-256. |
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 |
投稿时间:2014-12-09 |
DOI: |
中文关键词: 丙型肝炎病毒 JFH1株 适应性突变 全长cDNA |
英文关键词:Hepatitis C Virus(HCV),JFH1 strain,adaptive mutation,full-length complementary DNA |
基金项目:深圳市科技计划项目(201302096);深圳市新发传染病重点专科基金. |
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中文摘要: |
目的 构建丙型肝炎病毒JFH1全长cDNA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pBlueScript2ks(+)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cDNA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。 |
英文摘要: |
Objective To construct the full length complementary DNAs of HCV JFH1 strains with adaptive mutations. Methods Three HCV gene fragments (1.5 kb,4.1Kb and 4.1kb) 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 JFH1'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 JFH1. |
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