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用ＨｐａⅠ和Ｂｇ１Ⅱ双酶切３２ｋｂＨＢＶＤＮＡ，得到近１１ｋｂ大小的完整ＨＢｘ基因，利用绿豆芽核酸酶将ＨＢｘ基因、载体ｐＢＲ３２２的ＢａｍＨⅠ酶切粘性末端切成平末端，在Ｔ４ＤＮＡ连接酶作用下进行体外重组后转化到大肠杆菌中，获得亚克隆Ｅ．ｃｏｌｉＪＭ１０９ｐＢＲ３２２ＨＢｘ，并经药物平板筛选、电泳分析和ＤＩＧ标记的探针做Ｓｏｕｔｈｅｒｎ印迹杂交实验证实。Ｅ．ｃｏｌｉＪＭ１０９ｐＢＲ３２２ＨＢｘ亚克隆的建立对进一步研究ＨＢｘ基因的功能、肝癌发生的机理以及乙型肝炎病毒感染的诊断等均有重要意义
Hpa Ⅰ and Bg1 Ⅱ double enzyme digestion 3 2kbHBVDNA to get the size of the complete 1 1kb HBx gene, the use of mung bean sprouting nuclease HBx gene, vector pBR322 BamH Ⅰ enzyme cutting ends sticky ends cut in the T4 DNA ligase After in vitro recombination into E. coli, subclone E was obtained. coliJM109pBR322-HBx and confirmed by Southern blot hybridization with drug plate screening, electrophoresis and DIG-labeled probes. E. The establishment of subcellular cloning of JMJ910BR322-HBx is of great significance for the further study of the function of HBx gene, the mechanism of hepatocarcinogenesis and the diagnosis of hepatitis B virus infection