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董庆鸣, 何忠平, 庄辉, 宋淑静, 戴旺苏. C型乙型肝炎病毒的全基因组序列分析[J]. 中国公共卫生, 2003, 19(11): 1324-1325.
引用本文: 董庆鸣, 何忠平, 庄辉, 宋淑静, 戴旺苏. C型乙型肝炎病毒的全基因组序列分析[J]. 中国公共卫生, 2003, 19(11): 1324-1325.
DONG Qing-ming, HE Zhong-ping, ZHUANG Hui, . Complete genome sequence analysis of HBV genotype Cstrain[J]. Chinese Journal of Public Health, 2003, 19(11): 1324-1325.
Citation: DONG Qing-ming, HE Zhong-ping, ZHUANG Hui, . Complete genome sequence analysis of HBV genotype Cstrain[J]. Chinese Journal of Public Health, 2003, 19(11): 1324-1325.

C型乙型肝炎病毒的全基因组序列分析

Complete genome sequence analysis of HBV genotype Cstrain

  • 摘要:
      目的   探索乙型肝炎病毒(HBV)全基因组的结构特点.
      方法   应用直接PCR基因分型法确定HBV基因型, 然后随机选择1例单纯HBVC基因型感染的患者血清, 分3个相互重叠片段扩增HBV基因并分别克隆测序.利用VectorNTISuite7.0软件包、GeneDoc2.6和TreeView1.5, 将该3个基因片段拼接为全基因组序列, 命名为BDHB1, 并进行基因进化树分析和同源性比较.
      结果   HBVC基因型序列全长为3215bp, 其中A占222%, G占22.4%, C占26.8%, T占28.6%;有S、C、P和X区4个开放读码框架(ORFs); HBsAg亚型为adr; HBVRT区549~552aa为YMDD, 未发生变异; 未发现前C区nt1896G变异: BCP区nt1762、1764发生双突变.对HBV分段序列及全基因组序列进行基因进化树分析显示, BDHB1与HBVVC基因型的同源性最高.
      结论   BDHB1基因组符合HBVC基因型结构特点, 在BCP区存在双突变.分片段扩增再拼接可作为全基因组序列分析方法之一.

     

    Abstract:
      Objective   To investigate the characteristic of a complete HBV genome.
      Methods   The genotypes of HBV were determinated by PCR genoty ping, and the complete HBV genome was amplified through three over lapping segments amplification from the serum of a randomly selected patient infected with genotype C of HBV alone.The three amplicons were cloned into pGEM-T easy vector and sequenced.The sequences of the three amplicons were spliced as a complete HBV genome named as BDHB1, and then analyzed for phylogenetic tree and homologous analysis with Vector NTI Suite 7.0, GeneDoc 2.6 and TreeView1.5.
      Results   The genome of BDHB1 was 3215 bp long and the base A, G, C and T were 22.2%, 22.4%, 26.8% and 28.6%, respectively.It contained four over lapping open reading frames(ORFs): S, C, P and X region.Its HBsAg subtype was adr.There was no YMDD mutation in codons 549-552aa of RT region.No mutation occured at nt1896 in the precore region.A to T and G to A continuous mutations were detected atnt 1 762 and 1 764 in the BCP(Basic Core Promoter)region.Phylogenetic tree analysis of the overlapping segments and the complete genome both showed that BDHB1 was most homologous with HBV genotype C.
      Conclusion   The characteristic of BDHB1 was most similar to HBV genotype C.There were double mutations in BCP region of the strain.The overlapping segments amplification and splicing could be one of methods for the complete genome analysis.

     

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