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中文摘要：
目的建立血清标本经聚合酶链反应（PCR）扩增乙型肝炎病毒（HBV）全长基因的新方法，并初步进行克隆分析，从而为HBV分子生物学及致病机理研究奠定基础。方法针对位于整个HBV基因序列中的两个缺口区设计含酶切位点的引物，扩增3．2kbHBV全长DNA，经酶切及连接后克隆入PUC18载体。结果用PCR方法成功获得了3．2kbHBVDNA，经Sal Ⅰ酶切克隆人PUC18载体，获得重组质粒PUC18／HBV3．2，酶切鉴定和PCR扩增证实重组质粒中含有3．2kbHBV全长DNA，成功构建了HBV全基因克隆。结论成功建立了从患者血清中克隆HBV全基因组的方法，为从全基因水平深入研究乙型肝炎病毒变异与致病机理的关系奠定了基础。
英文摘要：
Objective To develop a new method for amplification and cloning of a full-length HBV genome by polymerase chain reaction （PCR）, and to lay the foundation for the HBV molecular biology and pathogenesis studies by preliminary cloning analysis. Methods A pair of primers containing restriction sites which were located at the nick region of the HBV molecule were used to amplify the 3.2 kb full-length HBV genome. PCR products were isolated and cloned into vector PUC18. Results The 3.2 kb full-length HBV genome was successfully obtained by this method. The recom- binant plasmid PUC18/HBV3.2 was constructed after the PCR products digested by restriction endonuelease Sal-Ⅰ and cloned into vector PUC18, identified with restricting endonuelease and PCR amplification, plasmid PUC18/HBV3.2 containing the 3.2 kb full-length HBV genome was successfully constructed. Conclusion The cloning of full-length HBV genom from serum samples is successfully established, which could serve to further study the relationship between HBV mutation and pathogenesis.
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