Construction and identification of recombinant lentiviral vector containing HIV-1 Tat gene and its expression in 293T cells

摘要

Objective:To construct a lentiviral vector expressing HIV-1 Tat and identify its expression in 293T cells. Methods:The gene fragment of HIV-1 Tat 101 was subcloned to lentiviral transfer vector pHAGE-CMV-MCS-IZsGreen,which was named pHAGE-Tat.Then the constructed pHAGE-Tat was used to co-transfect the packing 293T cells,together with the packaging plasmids pMD2.G and psPAX2.The packaged viral particles designated LV-Tat were used to infect the 293T cells and the viral titer was calculated.The expression of HIV-1 Tat in 293T cells was confirmed using RT-PCR and western blot.Results:The recombinant lentiviral vector was successfully constructed and could express HIV-1 Tat in 293T cells.The virus titer was 5.73×106 ifu/ml.Conclusion:The successfully constructed recombinant lentiviral vector makes a strong foundation for further exploring the possible role of HIV-1 Tat in the development of prostate cancer.