Aim: To construct a recombinant adenovirus vector containing p38MAPK gene and to identify its expression in mouse osteoblast-like cells, MC3T3-E1, in vitro.
Methods: The p38MAPK gene was amplified by PCR from mouse liver cDNA library, and inserted into pMD18-T vector and sequenced. Double digested with Bgl II and Hind III, p38MAPK gene was inserted into pShuttle-CMV. This recombinant plasmid was linearized by Pme I and electronically transfected into BJ5183 to get the recombinant adenovirus vector Ad-p38MAPK. Then the Ad-p38MAPK was obtained by packaging Pac I linearized in AD293 cells. The recombinant adenovirus expressing p38MAPK was infected into osteoblast- like cells(MC3T3-E1).The expression of exogenous p38MAPK was observed by Western blot.
Results: The recombinant plasmid Ad-p38MAPK was successfully generated, which increased p38MAPK protein expression levels in MC3T3-E1.
Conclusion: The bioactive recombinant adenovirus Ad-p38MAPK has been successfully constructed. The study laid a foundation for further research of the function of p38MAPK in osteoblast.