Construction of eukaryotic expression vector pcDNA3.1-bFGF and expression of the vector in canine bone marrow stem cells

Objective To explore the expression of eukaryotic expression vector bFGF in canine bone marrow stem cells(BMSCs) and to further study the application of bFGF gene transfer in bone tissue engineering.Methods Total cellular RNA of human glioma BT325 was isolated and RT-PCR amplification of the cDNA fragments of bFGF was performed.The fragment was cloned into vector pcDNA3.1.The eukaryotic expression vector was recombined and then transfected by lipofectamin into canine BMSCs.bFGF expression was analyzed using Western blot and RT-PCR.Results The recombinant vector pcDNA3.1-bFGF was identified by restriction enzyme analysis,PCR amplification and nucleotide sequencing.bFGF was successfully expressed in canine BMSCs.Conclusion The eukaryotic expression vector pcDNA3.1-bFGF,which can be expressed in canine BMSCs,has been successfully constructed.