Construction and identification of human FGF-9 lentiviral expression vector

Objective To construct a lentiviral vector carrying human FGF-9 gene, detect its expression, and pack the lentiviral vector, in order to provide the basis for studying the function of FGF- 9 in vivo and in vitro and its relationship with tumor. Methods Full-length of FGF-9 gene was amplified by PCR, and cloned into the plasmid pGC-FU-3FLAG-SV40-EGFP, PCR and gene sequencing were used to identify positive clones. 293T cells were transfected by the recombinant plasmid pGC-FU-3FLAG-SV40-EGFP, and a fluorescence microscope was used to observe the expression of eGFP, and Western-blot was used to detect the expression of FGF-9. 293T cells were then co-transfected with recombinant plasmid pGCFU-3FLAG-SV40-EGFP, pHelper 1.0 and pHelper 2.0, and the titer of lentivirus was determined by RT-qPCR. Results The recombinant expression vector was successfully constructed, eGFP and FGF-9 can express successfully in 293T cells, and lentiviruses were successfully packed by the 293T cells, the titer was 2×108TU/mL. Conclusion The recombinant expression vector is successfully constructed, and lentiviruses are successfully packed, it's the foundation for further exploration on FGF-9and its relationship with tumor.