Expression And Identification Of Recombinant Plasmid Of PSecTag2/B-OPG Into Beagle Dog Bone Marrow Stromal Cells In Vitro

Objective: In odder to treatment periodontitis by using tissue engineering and gene engineering technology, we establish transiently expression system of BMSCs modified by opg gene and detect its expression by using eukaryotic secreted expression pSecTag2/B-OPG plasmid.Method: Adult Beagle dog bone marrow about 2-3ml were drawled by puncture.Cells were isolated by a whole bone marrow adherence. After expanding and culture 3 passages, the identified recombined plasmid was transiently transfected into BMSCs by Lipofectamine 2000 and OPG expression in BMSCs was determined by RT-PCR and Western-blot.Results: The fragments of the recombinant plasmid digested with Hindâ…¢,EcoR I and BamHâ… and examined by 10g/L agarose electrophoresis were consistent with predicted size. The sequence of opg was identical to the sequence provided by GenBank [gi:33878056]. opg gene transcribing in Basks was confirmed by RT-PCR and OPG sustainable expressing in BMSCs was confirmed by Western-blot in 39 days. Conclusion: The transiently expression system of BMSCs modified by opg gene was successfully established.