Objective: To construct a mammalian expression vector of basic fibroblast growth factor (bFGF) and to investigate the expression of bFGF in vitro and in vivo.
Methods: A mammalian expression vector pcDNA3. 1/ myc-His(-)C-bFGF was constructed with gene cloning technique. The mammalian expression system was prepared and purified. The expression of bFGF cDNA in cultured transfected cells was examined by RT-PCR and cell immunohistochemistry. The recombinant plasmids, pcDNA3. 1/myc-His(-)C-bFGF and pCD2-VEGF121, were transferred into rabbit cervical muscle by direct injection of plasmid following electric pulses in vivo. The transferred gene expression and the biological effect were measured by use of histochemistry and immunohistochemistry analysis.
Results: The eukaryon expression system pcDNA3. 1/myc-His (-)C-bFGF could express the target protein bFGF in vitro. The recombinant plasmids, pcDNA3. 1/myc-His(-)C-bFGF and pCD2-VEGF121 were transferred into muscles flap in vivo successfully. The active proteins bFGF and VEGF121 were expressed at high levels. Blood vessels increased significantly in the muscles, and blood circulation was improved by local angiogenesis.
Conclusion: The eukaryon expression vector of bFGF is constructed and can be expressed successfully in vitro and in vivo. That is a primary preparation for the research on tissue transplantation and tissue engineering with bFGF gene therapy.