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  • ISSN 1674-8301
  • CN 32-1810/R
Volume 24 Issue 1
Dec.  2009
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Bingbing Wei, Ninghan Feng, Feng Zhou, Chun Lu, Jiantang Su, Lixin Hua. Construction and identification of recombinant lentiviral vector containing HIV-1 Tat gene and its expression in 293T cells[J]. The Journal of Biomedical Research, 2010, 24(1): 58-63.
Citation: Bingbing Wei, Ninghan Feng, Feng Zhou, Chun Lu, Jiantang Su, Lixin Hua. Construction and identification of recombinant lentiviral vector containing HIV-1 Tat gene and its expression in 293T cells[J]. The Journal of Biomedical Research, 2010, 24(1): 58-63.
shu

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

  • 1.

    Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China

  • 2.

    Department of Microbiology and Immunology, Nanjing Medical University, Nanjing 210029, China

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☆This study was supported by China Postdoctoral Science Foundation funded project (20080441064).

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    • Abstract
  • 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 5.73×106 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.
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