人免疫缺陷病毒假病毒药物筛选模型的构建及其应用  

Construction and Application of Pseudotyped Lentivirus Cell Model for Screening Anti-HIV Drug

作　　者：李建彬[1,2] 陈斌[1] 米志强[1] 安小平[1] 李存[1] 刘大斌[1] 王晓娜[1] 范华昊[1] 方祥[2] 童贻刚[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071 [2]华南农业大学食品学院,广东广州510642

出　　处：《生物技术通讯》2012年第4期481-484,共4页Letters in Biotechnology

基　　金：国家自然科学基金(30872223);国家科技重大专项课题(2008ZX10001-013);病原微生物生物安全国家重点实验室开放课题(PBS2009A-04)

摘　　要：目的:构建人免疫缺陷病毒(HIV)假病毒模型,用多种HIV逆转录酶和蛋白酶抑制剂作用于该模型,以检测其是否能有效用于HIV抑制药物的筛选。方法:通过载体改造获得最终慢病毒载体puc18-NL4-3-LUC-stop,其中含有萤光素酶基因,将该载体与包膜质粒VSV-G共转染293FT细胞,包装产生HIV假病毒,在假病毒包装和病毒感染293FT细胞的过程中加入蛋白酶和逆转录酶抑制剂,通过检测感染细胞中萤光素酶的表达来检测该模型的有效性,并利用此模型检测药物的抗病毒效果。结果:将HIV逆转录酶和蛋白酶抑制剂作用于该假病毒模型时发现萤光素酶的表达得到很大程度的抑制。结论:建立了HIV假病毒药物筛选模型,该模型以萤光素酶基因作为报告基因,快速灵敏,在抗HIV药物筛选中有一定的应用价值。Objective： To evaluate the effectiveness of a VSV-G pseudotyped lentivirus cell model for screening anti-HIV（human immunodefieiency virus） drug. Methods： The vector puelS-NL4-3-LUC-stop containing luciferase reporter gene was constructed and co-transfected into the 293FT cells with plasmid encoding vesicular stomatitis virus glycoprotein to produce VSV-G pseudotyped lentivirus. HIV protease and reverse transcriptase inhibitors were added in the process of lentivirus production and infection of 293FT cells. Expression of luciferase was de- tected to verify the validity of the model. The cell model was used to test the effectiveness of anti-HIV drugs. Resuits： When using protease and reverse transeriptase inhibitors in this model, we found that the expression of luciferase decreased dramatically compared with control. Conclusion： This model can be applied to screen anti-HIV drug quickly and accurately with luciferase gene as reporter.

关 键 词：人免疫缺陷病毒 慢病毒载体 假病毒 人免疫缺陷病毒抑制剂 药物筛选 

分 类 号：R373[医药卫生—病原生物学] Q78[医药卫生—基础医学]