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 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 5.73×10 6ifu/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.
Highlights
Prostate cancer is one of the most common malignant diseases in men[1]
The oncoviruses that predispose infected individuals to be more vulnerable to cancer include Kaposi's sarcoma-associated herpesvirus (KSHV)[2,3,4], Epstein–Barr virus, [5,6] and human papilloma virus[7,8,9], human T-lymphotropic virus[10,11] and hepatitis virus[12,13,14]
The insert of HIV-1 transcription protein (Tat) was confirmed by PCR, double-enzyme digestion (Fig. 1) and gene sequencing. pHAGE-Tat, pMD2.G and psPAX2 were used to co-transfect the 293T cells, and the viral particles were collected 48 and 72 hours later
Summary
Prostate cancer is one of the most common malignant diseases in men[1]. The mechanism of prostatic carcinogenesis and development is still unclear. The oncoviruses that predispose infected individuals to be more vulnerable to cancer include Kaposi's sarcoma-associated herpesvirus (KSHV)[2,3,4], Epstein–Barr virus , [5,6] and human papilloma virus[7,8,9], human T-lymphotropic virus[10,11] and hepatitis virus[12,13,14]. KSHV, known as herpesvirus-8 (HHV-8), is a Because of the increasing number of human beings infected with KSHV and HIV-1, it has become one of the most commonly studied oncoviruses and has attracted our attention.
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