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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 construct human immunodeficiency virus (HIV) pseudovirions model and use a variety of HIV reverse transcriptase and protease inhibitors in this model to test whether it can be effectively used in the screening of HIV inhibitory drugs. METHODS: The final lentiviral vector, puc18-NL4-3-LUC-stop, was constructed by vector transformation and contained the luciferase gene. The vector was cotransfected into 293FT cells with the envelope plasmid VSV-G and packaged into HIV pseudovirus. The protease and reverse transcriptase inhibitors were added during the process of pseudowire packaging and virus infecting 293FT cells. The efficiency of this model was tested by detecting luciferase expression in infected cells. The model was used to detect the antiviral effect of the drug . Results: When HIV reverse transcriptase and protease inhibitors were administered to the pseudovirion model, luciferase expression was found to be largely inhibited. CONCLUSION: A model of HIV pseudovirus screening is established. The luciferase gene is a rapid and sensitive reporter gene and has certain value in the screening of anti-HIV drugs.