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Specific Marking of HIV-1 Positive Cells using a Rev-dependent Lentiviral Vector Expressing the Green Fluorescent Protein
Published on: September 23, 2010
Construction and identification of recombinant lentiviral vector containing HIV-1 Tat gene and its expression in 293T
Bingbing Wei1, Ninghan Feng, Feng Zhou
1Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Objective:
To construct a lentiviral vector expressing HIV-1 Tat and identify its expression in 293T cells.
Methods:
The gene fragment of HIV-1 Tat101 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 5.73×10(6) 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.
