Construction and expression of pLenti6/v5-R-K with CC-intrakine
AIM: To study the HIV-based lentiviral vector, pLenti6/V5-R-K, which contained the gene CC-intrakine (RAN-TES-K) and to provide a basis for the gene therapy of HIV-1 infection. METHODS: A pLenti-based expression vector, pLen-ti6/V5-D-TOPO?, was used to produce the lentiviral vector, which was cotransfccted with the ViraPowerTM Packaging Mix (pLP1, pLP2, and pLP/VSVG) into 293 FT cells to produce a replication-incompetent lentivius stock. After titrating the lentiviral stock using HeLa cells, the expression of the interest gene of RANTES was investigated by indirect immumofluorescence in transduced HeLa cells. RESULTS: The lentiviral expression vector, pLenti6/V5-R-K, was confirmed by enzymatic digestion and sequencing. The lentivirus stock was constructed in 293 FT cell line. The fluorescein was mainly scattered in cytoplasm and amph-nucleus in transduced HeLa cells. CONCLUSION: These findings demonstrate the ability of the lentiviral vector to transduce multiple genes into HeLa cells and its potential therapeutic roles in the treatment of HIV-1 infection.
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