title: [Construction of the recombinant plasmid of pcDNA3.0-RKIP and expression in PC12 cells].
abstract: OBJECTIVE
  To construct the eukaryotic expression vectors of RKIP plasmid and detect its expression in PC12 cells.
  
  
  METHODS
  The coding sequence of RKIP was generated by nested-PCR using total RNA extracted from the root ganglion neurons of rats. RKIP gene was cloned into the eukaryotic expression vector pcDNA3.0. After restriction enzyme analysis and sequence identification, the recombinant plasmid was transfected into PC12 cells with non-liposome mediated method by Vigofect. The expression of RKIP was detected by Western blot.
  
  
  RESULTS
  The results of enzyme analysis and sequencing both identified DNA sequence of recombinant plasmid pcDNA3.0-RKIP correctly. The expression of RKIP increased obviously after transfection into PC12 cells.
  
  
  CONCLUSIONS
  The eukaryotic expression plasmid of pcDNA3.0-RKIP was constructed successfully and it can be sustainly expressed in PC12 cells. This provides experimental basis for further study on the neurological function of RKIP.
authors.affiliation: Institute of Radiation Medicine, Academy of Military Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China. shanialin@126.com
authors.forename: Tao
authors.initials: T
authors.lastname: Lin
authors.forename: Hong-yan
authors.initials: HY
authors.lastname: Zuo
authors.forename: De-wen
authors.initials: DW
authors.lastname: Wang
authors.forename: Rui-yun
authors.initials: RY
authors.lastname: Peng
authors.forename: Pei-lan
authors.initials: PL
authors.lastname: Zhou
authors.forename: Shui-ming
authors.initials: SM
authors.lastname: Wang
authors.forename: Ya-bing
authors.initials: YB
authors.lastname: Gao
authors.forename: Shao-xia
authors.initials: SX
authors.lastname: Wang
mesh_terms: D000818:Animals; D016716:PC12 Cells; D049488:Phosphatidylethanolamine Binding Protein / Q000235:genetics*; D010957:Plasmids; D051381:Rats; D011995:Recombination, Genetic; D014162:Transfection
chemical_list: C550693:PEBP1 protein, rat; D049488:Phosphatidylethanolamine Binding Protein