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《Chinese Journal of Cellular and Molecular Immunology》 2011-04
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The construction of recombinant adenovirus containing p38MAPK gene and its expression in mouse osteoblast-like cells(MC3T3-E1)

BAI Xue-dong1,HU Yun-yu2,LI Hang2,HOU Li-sheng1,LI Hai-feng1 1Department of Orthopaedics,the Naval General Hospital,Beijing 100037;2Xijing Hospital,Fourth Military Medical University,Xi'an 710032,China  
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 BglⅡ 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.
【Fund】: 国家自然科学基金资助项目(30772261)
【CateGory Index】: R329
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