Construction of a recombinant retroviral vector containing human bFGF cDNA and its expression in mammalian cells

Objective To constract a retroviral mediated expression system of human basic fibroblast growth factor (bFGF). Methods Recombinant retroviral vecter pLXSN bFGF was generated by cloning a full length human bFGF cDNA into retroviral vecter pLXSN. COS 7 cells were infected with the viral supernatant from the highest productive PA317 clones. RT PCR was used for the detection of bFGF mRNA and bFGF protein in the COS 7 cells was detected with Western Blotting. Results The restriction enzyme analysis with EcoR Ⅰ and Xho Ⅰ showed that the recombinant retroviral vector had been constructed correctly. The titer assayed on NIH3T3 cells was up to 2.3×10 4 CFU/ml. Using the RT PCR and Western Blotting methods, the bFGF mRNA and bFGF protein were expressed in COS 7 cells after pLXSN bFGF infection. Conclusion The constructed retroviral vector is able to generate effective expression of human bFGF in mammalian cells, with potential utility in the gene therapy for central nervous system diseases such as Parkinsonism dimentia.