title: [The construction of recombinant adenovirus containing p38MAPK gene and its expression in mouse osteoblast-like cells (MC3T3-E1)].
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Orthopaedics, the Naval General Hospital, Beijing 100037, China. baixuedonghx@yahoo.com.cn
authors.forename: Xue-dong
authors.initials: XD
authors.lastname: Bai
authors.forename: Yun-yu
authors.initials: YY
authors.lastname: Hu
authors.forename: Hang
authors.initials: H
authors.lastname: Li
authors.forename: Li-sheng
authors.initials: LS
authors.lastname: Hou
authors.forename: Hai-feng
authors.initials: HF
authors.lastname: Li
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D051379:Mice; D010006:Osteoblasts / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics*
chemical_list: D048051:p38 Mitogen-Activated Protein Kinases