构建不同筛选特性的HBV X基因真核表达载体  

Construction of Two Eukaryotic Expression Vectors of HBV X Gene with Different Selection Characteristics

作　　者：贺兴鄂[1] 雷建华[1] 杨旭[1] 王文龙[1] 罗红雨[1] 梁骏[1] 

机构地区：[1]中南大学湘雅二医院肝病研究中心,湖南长沙410011

出　　处：《中西医结合肝病杂志》2005年第5期274-276,共3页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金(No.30371402)

摘　　要：目的:构建不同筛选特性的两个HBV X基因真核表达载体。方法:从质粒pEcob6中PCR扩增HBV X全基因,利用pCEP4和pcDNA3.1(+)两者多克隆位点的特点,选用pGEMR○-T Easy Vector构建中间载体pEasy-X,分别酶切后连接特异性片段构建载体pCEP4-X和pcDNA3.1(+)-X。结果:从质粒pEcob6成功PCR扩增出HBV X全基因并克隆至质粒pEasy-X,酶切、PCR及测序均证实真核表达载体质粒pCEP4-X和pcDNA3.1(+)-X构建成功。结论:具有不同筛选特性的两个HBV X基因真核表达载体业已成功构建。Objective： To construct two eukaryotic expression vectors of HBV X gene with different selection characteristics. Methods： X gene from plasmid pEcob6 was amplified by PCR assay with specific primers. With the characteristics of multiple clone sites in pCEP4 and pcDNA3.1 （ ＋ ）, pGEM-T Easy Vector was selected to construct intermediate vector pEasy-X. After enzyme digestion, specific fragments were linked to construct pCEP4-X and pcDNA3.1 （ ＋ ） -X vectors. Results： X gene was succeesfully cloned from pEcob6 by PCR assay and the amplified gene products were successfully inserted into pEasy-X vectors. The successful insertion of HBV X gene into pCEP4-X and pcDNA3.1 （ ＋ ） -X vectors was confirmed by enzyme digestion, PCR assay and sequencing. Conclusion： Two eukaryotic expression vectors of HBV X gene with different selection characteristics were successfully constructed.

关 键 词：HBV X基因 真核表达载体 构建 基因真核表达载体 HBVX 筛选 PCDNA3.1(+) PCR扩增 载体质粒 特异性片段 多克隆 

分 类 号：R373.21[医药卫生—病原生物学]