The Establishment of HIV-1 Tat Protein Binding Tar Model for Screening HIV-1 Inhibitors

Objective To establish HIV-1 Tal binding to Tar model.Methods HIV-1 Tat protein eukaryotic expression plasmid(pCDNA3.1(+)-Tat)and luciferase report gene HIV-1 LTR-luc(LTR-LUC) were constructed and transfected into HeLa cells with lipofectamine 2000.The expression of HIV-1 Tat protein in HeLa cells was detected with Iuminometer.HIV-1 Tat protein binding tar model for screening HIV-1 inhibitors was thereby constructed.Results The abstractions of Xanthoceras sorbifolia and Perilla frutescens and so on were detected and DRB(5,6-dichloro-1-β-D-robofuranosylbenzimidazole) was positive control.The stability of the model was tested and regulated.Conclusion The repeated experiments indicate that the matching of purpose gene and report gene,the rate of new calf serum in medium during transfection,density of the transfected cells,condition of the untransfected cells and the action time of drugs affect the stability and sensitivity of the model.