The Expression And Antibacterial Activity Of Antimicrobial Peptides PR-39 In Rabbit Bone Mesenchymal Stem Cells Mediated By Lentiviral Vector

Objective : To construct the proline-arginine rich 39-amino acid peptide(PR-39) lentiviral vector, and introduce it into bone mesenchymal stem cells(BMSC), the expression and antibacterial activity of PR-39 was detected.Methods:The CDS sequence of PR-39 was amplified by PCR from pBluescript II SK-PR-39, and then the gene was recombined into the transfer plasmid pGC-FU. The three plasmids of lentiviral vector, transfer plasmid pGC-FU-PR-39, packaging plasmid pHelper1.0,pHelper2.0, which were cotransfected to 293T cells mediated by lipofectamin 2000 to produce lentiviral particles. The tite of viral particles was detected by real-time PCR. The rabbit BMSC were prepared, and then were infected by PR-39 lentiviral particles. The transfection efficiency was observed with fluorescent microscope and flow cytometry, and the expression of PR-39 was detected by RT-PCR, and the antibacterial activity also was investigated. Results:The tite of viral particles was 2×109 TU/ml, and it was showed transfection efficiency was 74.09%. PR-39 gene was expressed successfully in BMSC,and PR-39 inhibited the staphylococcus aureus significantly, and bacteriostatic rate was 98.43% (P<0.05).Conclusion:Antimicrobial peptides PR-39 is expressed successfully in rabbit BMSC mediated by lentiviral vector, and it shows significant antibacterial activity.