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目的：克隆中国人庚型肝炎病毒５′端基因，并对其ｃＤＮＡ序列进行分析。方法：应用改进的ｃＤＮＡ末端快速扩增法，即利用特异引物合成第一链ｃＤＮＡ，纯化后在其３′端加ｐｏｌｙ（Ａ）“多聚尾”，用Ｏｌｉｇｏｄ（Ｔ）－ａｎｃｈｏｒ等引物进行“巢式”ＰＣＲ扩增，从河北省固安县一献血员血清中获得ＨＧＶ５′端基因片段，与相应载体连接后转化宿主菌，随机挑选４个阳性克隆进行序列分析。结果：４个克隆的基因序列均有差异，且与３株美国ＨＧＶ的５′最末端都存在一定变异。结论：中国人庚型肝炎病毒５′端存在基因多样性，提示庚型肝炎病毒的５′非编码区并不是很保守。用此方法克隆单链ＲＮＡ病毒基因组的５′端，国内尚未见报道
Objective: To clone the 5 ’end gene of Chinese hepatitis G virus and analyze its cDNA sequence. METHODS: The first-strand cDNA was synthesized by using the rapid amplification of cDNA ends by using specific primers. After purification, poly (A) “poly tail” was added to the 3 ’end of the cDNA. The primers were used for Oligod (T) In the “nested” PCR amplification, a 5 ’end gene fragment of HGV was obtained from a blood donor serum of Gu’an County, Hebei Province. After being connected with the corresponding vector, the HGV 5’ end gene fragment was transformed into host bacteria. Four positive clones were randomly selected for sequence analysis. Results: The sequences of the four clones were different, and there was some variation with the 5 ’end of the three American HGV strains. Conclusion: The diversity of 5 ’end of Chinese hepatitis G virus indicates that the 5’ non-coding region of hepatitis G virus is not very conservative. Cloning single-stranded RNA virus genome by this method 5 ’end, has not been reported in China