在大肠杆菌中利用反义RNA抑制乙肝病毒（HBV）X基因的表达  

INHIBTION OF HBV X GENE EXPRESSION BY ITS ANTISENS RNA IN E coli

作　　者：杨建琪[1] 张锡元[1] 李文鑫[1] 张晓刚[1] 王平[1] 

机构地区：[1]武汉大学生命科学学院,中国预防医学科学院病毒所基因工程国家重点实验室

出　　处：《武汉大学学报（自然科学版）》1995年第4期475-481,共7页Journal of Wuhan University(Natural Science Edition)

摘　　要：将乙肝病毒（ＨＢＶ）ａｙｗ株完整的Ｘ基因正向重组到原核表达质粒ｐＢＶ－２２１的ＰＬ启动子下游，得到能表达Ｘ蛋白的重组质粒ｐＢＶ－ＨＢＶ（＋）；同时将Ｘ基因反向重组到原核表达质粒ｐＢＶ－２２０的ＰＬ启动子下游，得到能转录Ｘ基因反义ＲＮＡ的重组质粒ｐＢＶ－ＨＢＸ（－）．利用这两个质粒，构建出能同时转录Ｘ基因ｍＲＮＡ和反义ＲＮＡ的重组质粒ＰＥＸ．ＡＮ－ＨＢＸ，并在原核水平上，证实了反义ＲＮＡ对Ｘ基因的表达具有明显的抑制作用，从而为在真核水平上利用反义ＲＮＡ对Ｘ基因进行调控的研究提供了有利的基础．In both direction,placing the entire HBV X gene DNA sequence under thecontrol of PL promoter of expression plasmids pBV-221and PBV-220. The result showed thatthe recombined plasnlids pBV-HBX(+) and PBV-HBX(-) were constructed suuessfully. InPBV-HBX(+), HBV X gene was cloflctl in normal ORF (Open Reading Frame) and couldexpress correct X protein. But for pBV-HBX(- ),the X gene' S direction was opposite and itcould only transcript the antisense RNA of HBV X gene. With the two plasmids, plasmidPEX. AN-HBX which could transcript HBV X gene mRNA and antisense RNA at same timewas constructed.With the procaryotic expression system transformed with the above plasmids, HBV Xprotein was expressed successfully in E coli DH5α and obvious inhibition to this expressionby antisense RNA was ohservated in E coli DH5α.

关 键 词：RNA HBV X基因 乙型肝炎病毒 大肠杆菌 基因表达 

分 类 号：R512.62[医药卫生—内科学]