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凌斌华, 庄辉, 汪兴太, 李奎, 崔怡辉, 朱永红. 庚型肝炎病毒PCR产物直接序列测定方法的建立及应用[J]. 中国公共卫生, 1998, 14(3): 137-138.
引用本文: 凌斌华, 庄辉, 汪兴太, 李奎, 崔怡辉, 朱永红. 庚型肝炎病毒PCR产物直接序列测定方法的建立及应用[J]. 中国公共卫生, 1998, 14(3): 137-138.
Ling Binhua, . Establishment and Application of Direct Sequencing of Hepatitis G Virus PCR Products[J]. Chinese Journal of Public Health, 1998, 14(3): 137-138.
Citation: Ling Binhua, . Establishment and Application of Direct Sequencing of Hepatitis G Virus PCR Products[J]. Chinese Journal of Public Health, 1998, 14(3): 137-138.

庚型肝炎病毒PCR产物直接序列测定方法的建立及应用

Establishment and Application of Direct Sequencing of Hepatitis G Virus PCR Products

  • 摘要: 以PCR产物为测序模板,γ-32P-ATP标记的PCR扩增引物为测序引物,用Taq DNA聚合酶为测序酶,建立了庚型肝炎病毒(HGV)cDNA序列分析方法.应用本法测定1份PCR阳性产物的HGV cDNA序列,图谱呈清晰的序列梯,其序列与GBV-C(U36380)和HGV(U44402)的相应片段序列比较,同源性分别为85.65%(191/223)和85.2%(190/223).与用荧光法测定的另一株HGV序列有高度一致性.对同一样品两端测序,重叠部分序列完全一致.表明PCR产物直接测序法可用于HGV核酸序列分析.

     

    Abstract: Direct sequencing of hepatitis G virus(HGV)PCR products was established using the 32P end-labeled primer of PCR and Thermus aquat icus DNA polymerase(sequencing grade).By this sequencing strategy,HGV cDNA was sequenced.Clear sequence ladders were obtained and the sequence was highly identical with that of another HGV cDNA sequenced by the automatic laser fluorescent(ALF) DNA sequencer.The overlapped sequences were identical when one product was sequenced from both sides.In conclusion,direct sequencing of the PCR products can be used for nucleotide sequence analysis of HGV cDNA.

     

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