Apobec3B的真核表达及其抗HBV效应

目的：在真核细胞中表达Apobec3B,并探讨其在体外对HBV复制和表达的抑制作用。方法：应用RT-PCR法从人PBMC细胞中扩增Apobec3B基因,构建真核表达载体pcDNA3.1-Apo-bec3B,转染Hep G2 2.2.15细胞。W estern-blot鉴定细胞中Apobec3B蛋白的表达。ELISA法检测细胞培养上清液中HBsAg和HBeAg水平,RT-PCR检测HBV DNA和HBV mRNA水平。结果：经酶切及测序鉴定pcDNA3.1-Apobec3B成功构建,Apobec3B蛋白可在Hep G22.2.15细胞中成功表达。转染pcDNA3.1-Apobec3B重组质粒的Hep G2 2.2.15细胞的HBsAg、HBeAg、HBV DNA及HBVmRNA水平均明显低于转染空载体组的Hep G2 2.2.15细胞。结论：成功在真核细胞中表达Apo-bec3B,其可明显抑制了Hep G22.2.15细胞中HBV的复制与表达,为深入研究Apobec3B抗HBV的作用机制奠定基础。

Eukaryotic Expression of Apobec3B and the Effects of anti-HBV

Objective： To express Apobec3B in eukaryotic cells,and study the effects of Apobec3B on replication and expression of HBV in vitro.Method： The Apobec3B gene was extracted from PBMCs by RT-PCR.Eukaryotic expression vector pcDNA3.1-Apobec3B was constructed and transfected into Hep G2 2.2.15 cells.The expressional product was detected by Western-blot in Hep G2 2.2.15 cells.The levels of HBsAg and HBeAg were detected by ELISA.The levels of HBV DNA and HBV mRNA were determined by RT-PCR.Result： The pcDNA3.1-Apobec3B was constructed successfully by restriction analysis and sequence identification,Apobec3B was expressed efficiently in Hep G2 2.2.15 cells.The levels of HBsAg,HBeAg,HBV DNA and HBV mRNA in Hep G22.2.15 cells transfected pcDNA3.1-Apobec3B were significantly lower than that of Hep G2 2.2.15 cells which were transfected empty vectors.Conclusion： Apobec3B was expressed successfully in eukaryotic cells.It could obviously inhibit replication and expression of HBV in vitro.It is a basic work for the following study of the mechanism of anti-HBV effect s by Apobec3B.