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Published on: July 30, 2018
[The construction of recombinant adenovirus containing p38MAPK gene and its expression in mouse osteoblast-like cells
Xue-dong Bai1, Yun-yu Hu, Hang Li
1Department of Orthopaedics, the Naval General Hospital, Beijing 100037, China. baixuedonghx@yahoo.com.cn
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.
