摘要
目的观察小鼠体内乙型肝炎病毒(HBV)X蛋白(HBx)对HBV基因转录和复制水平的影响。方法应用高压注射体内转染法将野生型HBV重组质粒、HBx表达缺失的HBV重组质粒及HBx表达质粒DNA通过尾静脉快速注射入小鼠体内,应用Southern和Northern印迹分别检测小鼠肝组织中HBV DNA复制中间体及HBV mRNA水平。结果野生型及HBx表达缺失的HBV重组质粒注射组小鼠肝组织中均检测到HBV的复制与转录;HBx表达缺失的HBV重组质粒注射组HBV基因转录和复制水平较野生型HBV重组质粒注射组、HBx表达缺失HBV重组质粒与HBx表达质粒共注射组弱,生理盐水注射组未检测到HBV的转录与复制。结论HBx的表达缺失可导致体内HBV基因转录和复制水平的下降,而外源HBx的表达可逆转这种因HBx表达缺失引起的HBV复制及转录的减弱,提示HBx增强小鼠体内HBV基因转录和复制。
Objective To investigate the effects of HBx in regulating HBV transcription and replication in vivo.Methods Mice were rapidly injected with a large volume of wild-type or HBx-minus HBV genome and HBx expression plasmid DNA via tail vein. The transcription level of HBV mRNA was analyzed by Northern blot hybridization, and the levels of HBV DNA replication intermidiates were analyzed by Sounthern blot hybridization analysis. Results The HBV DNA replication intermidiates and RNA transcripts were detected in both wild-type or HBx-minus HBV genome injected groups, however were not detected in Sodium Chloride transfected group .The levers of HBV DNA replication intermidiates and RNA transcripts of HBx-minus HBV genome injected group were lower than wild-type HBV genome and HBx-minus HBV genome plus HBx injected groups. Conclusion The absence of HBx expression causes the reduction of HBV transcription and replication and the decreases could be restored by expression of HBx. The results suggested that HBx enhances HBV transcription and replication in vivo of mice.
出处
《四川医学》
CAS
2008年第6期629-631,共3页
Sichuan Medical Journal
基金
国家自然科学基金资助项目(No.30570064)
四川省应用基础研究项目(2006J13-087)
