Co-tranfection PR-39and BMP-9in C3H10T1/2Cells Mediated By Adenovirus Vector

Abstract： Objective: Construct a recombinant adenovirus vectorcontaining red fluorescent protein (RFP) gene and Proline-arginine rich39-amino acid peptide. The adenovirus vector containing red fluorescentprotein (RFP) gene and BMP-9was amplified。Introduce them intoC3H10T1/2cells, and then detect the expression and activity of PR-39andBMP-9.Methods: PR-39gene was amplified using PCR and inserted intopAdTrack-TO4vector． The pAdTrack-TO4-PR-39was confirmed byrestriction endonuclease digestion and gene sequencing, and thenpAdTrack-TO4-PR-39was transfected into competent AdEasier cells afterlinearized by PmeI to acquire recombinant adenovirus plasmid pAdPR-39.Then pAdPR-39was transfected into HEK293to acquire recombinantadenovirus(AdPR-39). Then AdPR-39and AdBMP-9were amplified.Mesenchymal stem cells C3H10T1/2will be infected by adenovirus, andthen transfected cells were examined under fluorescence field、RT-PCRand western blot detected restructuring adenovirus expression and theantibacterial activity also was investigated.Results: The pAdTrack-TO4-PR-39and pAdPR-39were constructed successfully. AdPR-39was successfully packaged in HEK293cells.AdPR-39and AdBMP-9could infect C3H10T1/2cells and express PR-39and BMP-9gene. staphylococcus aureus can be inhibited by the PR-39，and antibacterial rate was96.1%Conclusion: A recombinant adenovirus AdPR-39containing redfluorescent protein (RFP) reporter gene was successfully constructed. Theadenovirus AdPR-39and AdBMP-9could infect C3H10T1/2and expressPR-39and BMP-9gene. and it shows significant antibacterial activity.