title: [Construction of different mutants of HA-tagged human RAGE gene and their eukaryotic expression].
abstract: OBJECTIVE
  To construct eukaryotic expression vectors for HA-tagged receptor for advanced glycation end products (RAGE) mutants.
  
  
  METHODS
  Site-directed mutagenesis was applied to wild-type RAGE gene cloned in the pcDNA3 vector with HA tag to obtain the mutants pcDNA3-HA-RAGE(S391A), pcDNA3-HA-RAGE(S399A), pcDNA3-HA-RAGE(S400A), and pcDNA3-HA-RAGE(T401A). After identification by sequencing, the mutants were transfected into HEK293 cells, and the expression of these mutants were detected by Western blotting using anti-HA antibody.
  
  
  RESULTS
  The HA-tagged RAGE mutants constructed were verified successfully by sequencing, and highly expressed in HEK293 cells.
  
  
  CONCLUSIONS
  The success in constructing HA-tagged RAGE mutants, which are highly expressed in eukaryotic cells, may facilitate the functional study of RAGE in cell signal transduction.
authors.affiliation: Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China. chengweiwei1984@163.com
authors.forename: Wei-wei
authors.initials: WW
authors.lastname: Cheng
authors.forename: Yu-sheng
authors.initials: YS
authors.lastname: Li
authors.forename: Xiao-wei
authors.initials: XW
authors.lastname: Gong
authors.forename: Lin-lin
authors.initials: LL
authors.lastname: Zhao
authors.forename: Ji-gang
authors.initials: JG
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Deng
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
mesh_terms: D002460:Cell Line; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic