title: [Vector construction for the intracellular domain of human receptor for advanced glycation end- products and expression of the fusion protein].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector for GST-tagged intracellular domain of human receptor for advanced glycation end-products (hRAGE), and to study the function of the expressed fusion protein and identify its interacting proteins.
  
  
  METHODS
  The coding sequence of the intracellular fragment of hRAGE was amplified by PCR and inserted into pGEX-KG vector, a general GST fusion protein expression vector. After PCR identification, endonuclease digestion and DNA sequencing, the recombinant was transformed into E.coli BL21 to achieve the expression of GST fusion protein induced by isopropyl-beta-D-thiogalactoside (IPTG), followed by purification of the protein on glutathione-superflow resin.
  
  
  RESULTS
  The recombinant of GST/hRAGE-C was constructed and identified by PCR, endonucleases digestion and DNA sequencing. After protein expression was achieved in E. coli, a molecular mass of 35 kD GST fusion protein was purified, whose molecular mass and purity were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
  
  
  CONCLUSIONS
  The expression vector for intracellular domain of hRAGE has been successfully constructed and efficient expression of the fusion protein is obtained, which can be of value for further studies.
authors.affiliation: Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China. yjiang@263.net
authors.forename: Shan-chao
authors.initials: SC
authors.lastname: Zhao
authors.forename: Jing-hua
authors.initials: JH
authors.lastname: Liu
authors.forename: Zhi-jie
authors.initials: ZJ
authors.lastname: Li
authors.forename: Qing-he
authors.initials: QH
authors.lastname: Qin
authors.forename: Peng
authors.initials: P
authors.lastname: Deng
authors.forename: Fan-fan
authors.initials: FF
authors.lastname: Hou
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
mesh_terms: D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase