中文摘要：

目的：构建固有免疫分子TRIM5α的真核表达质粒,研究TRIM5α对HBV复制的影响。方法：通过巢氏RT-PCR技术从恒河猴肺组织中扩增出TRIM5α的编码基因,并将其克隆入pcDNA3.1真核表达载体。将TRIM5α真核表达质粒转染HepG2细胞,用Western blot的方法鉴定TRIM5α蛋白表达情况。将TRIM5α真核表达质粒与复制型HBV质粒通过磷酸钙沉淀法共转染HepG2细胞、通过尾静脉高压注射法共转染BALB/c小鼠。ELISA检测转染细胞上清及小鼠血清中的HBsAg和HBeAg;免疫组化检测小鼠肝组织HBcAg的表达;Southern blot检测转染细胞中HBV复制中间体。将TRIM5α真核表达质粒转染293T细胞3小时后,再将基于HIV-1结构的慢病毒载体三质粒系统转染293T细胞,通过检测转染细胞上清中的HIV-1 p24抗原含量来鉴定TRIM5α抗HIV-1功能。结果：成功构建TRIM5α真核表达质粒;过表达TRIM5α不能有效降低HBV抗原和复制中间体水平,但可抑制HIV-1 p24抗原的表达。结论：TRIM5α不能有效抑制HBV的复制。

英文摘要：

Objective：To construct a eukaryotic expression vector for TRIM5α and investigate whether TRIM5α have the ability to inhibit Hepatitis B virus （HBV） replication. Methods：The DNA fragment encoding TRIM5α was obtained from Rhesus monkey lung tissue by nested RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1. TRIM5α expression plasmid and HBV replication competent plasmid were co-transfected into HepG2 cells using the calcium phosphate precipitation method or co-delivered to mouse liver by hydrodynamic injection. ELISA was used to determine the levels of HBsAg and HBeAg, HBV core protein was visualized by immunohistochemical staining of liver tissues, and the HBV DNA replicative intermediates was detected by Southern blot. The anti-HIV ability of TRIM5α was evaluated by measuring the HIV-1 p24 level in culture supernatants. Results： The TRIM5α eukaryotic expression plasmid was successfully constructed. Overexpression of TRIM5α failed to decrease the expression level of HBV protein and replicative intermediates, but could decrease the HIV- 1 p24 level significantly. Conchusion：TRIM5α can＇t inhibit the replication of HBV.