title: [Eukaryotic expression and application of human CD226 Ig fusion protein containing 3C protease-restricted site].
abstract: OBJECTIVE
  To construct, express and characterize the eukaryotic expression vector of encoding human CD226 (PTA1) extracellular region Ig fusion protein gene containing 3C protease-restricted site.
  
  
  METHODS
  The gene fragment encoding extracellular region of human CD226 was cloned into the eukaryotic expression vector p-3C-Ig containing 3C protease-restricted site and human Ig Fc fragment gene. After sequencing, the vector was transfected into COS7 cells, and the expressed molecule was purified by affinity chromatography. Finally, the product was characterized by immunofluorescent staining and 3C protease digestion.
  
  
  RESULTS
  After expression and purification, the Ig fusion protein could bind effectively to the CD226 ligand expressed on ECV304 cells. The Fc fragment could also be cut off by 3C protease, so that the CD226 extracellular fragment was obtained.
  
  
  CONCLUSIONS
  The extracellular region of human CD226 Ig fusion protein with 3C protease-restricted site is expressed successfully in COS-7 cells, which lays the foundation for the structural and functional study of this molecule.
authors.affiliation: Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Xin-Hai
authors.initials: XH
authors.lastname: Zhang
authors.forename: Kun
authors.initials: K
authors.lastname: Yang
authors.forename: Wei
authors.initials: W
authors.lastname: Jia
authors.forename: Wei-Ming
authors.initials: WM
authors.lastname: Ouyang
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiao-Guang
authors.initials: XG
authors.lastname: Xu
authors.forename: Qi
authors.initials: Q
authors.lastname: Li
authors.forename: Bo-Quan
authors.initials: BQ
authors.lastname: Jin
mesh_terms: D000818:Animals; D019556:COS Cells; D056890:Eukaryota / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection
chemical_list: D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins