乙型肝炎病毒X基因在大肠杆菌中亚克隆与鉴定  

Subcloning and Identification of Hepatitis B Virus Gene in E.coli

作　　者：孙峥嵘[1,2,3] 鲁润铭 刘桂荣[1,2,3] 王桂珍 姚振宇[1,2,3] 佟宝军[1,2,3] 

机构地区：[1]沈阳市第六人民医院中心实验室 [2]中国医科大学微生物学教研室 [3]沈阳市公安局刑警支队技术科

出　　处：《中国公共卫生学报》1998年第5期280-281,共2页

摘　　要：用ＨｐａⅠ和Ｂｇ１Ⅱ双酶切３２ｋｂＨＢＶＤＮＡ，得到近１１ｋｂ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．ｃｏｌｉＪＭ１０９ｐＢＲ３２２ＨＢｘ，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．ｃｏｌｉＪＭ１０９ｐＢＲ３２２ＨＢｘ亚克隆的建立对进一步研究ＨＢｘ基因的功能。A complete HBx gene was obtained from the HBV DNA(3 2kb)digested with Bg1Ⅱ and Hpa Ⅰ.The cohesive terminal on HBx gene and vector pBR322 yielded by BamH Ⅰ were modified into blund ends with Mung Bean nuclease,then the two fragments were recombinated with T4DNA ligase in vitro.A subclonal E.coli JM 109 pBR322HBx was gained after transformation of the recombinant into E.coli,and was confirmed by drug plated selection、agarose electrophoresis and DIG Southern Blot.The establishment of subclonal E.coli JM 109 pBR322HBx is of great importance for the further research of HBx gene function,the mechanism of liver cancer and the diagnosis of hepatitis.

关 键 词：乙型肝炎病毒 HBX基因 SOUTHERN 

分 类 号：R446.112[医药卫生—诊断学]