title: [Construction and expression of green fluorescent protein fusion vector of p38 regulated/activated protein kinase].
abstract: OBJECTIVE
  To construct the expression vector of p38 regulated/activated protein kinase (PRAK) fused to green fluorescent protein which is capable of expression in mammalian cells.
  
  
  METHODS
  PRAK with His-tag in pET-14b expression vector was subcloned into the green fluorescent protein vector pEGFP-C2. The recombinant vector was then transfected into HeLa cells, followed by observation of the cells with fluorescent microscope.
  
  
  RESULTS
  Identification by enzyme digestion, PCR and sequencing confirmed successful construction of the recombinant vector, which was highly expressed in HeLa cells. Green fluorescence of the fusion protein EGFP-PRAK was observed mainly in the cell nuclei.
  
  
  CONCLUSIONS
  The expression vector of PRAK fused to green fluorescent protein is successfully constructed and expressed in mammalian cells, which may facilitate the study of intracellular localization and translocation of PRAK.
authors.affiliation: Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Peng
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Gong
authors.forename: Peng
authors.initials: P
authors.lastname: Deng
authors.forename: Qing-He
authors.initials: QH
authors.lastname: Qin
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
mesh_terms: D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D048051:p38 Mitogen-Activated Protein Kinases