title: [Construction of GPI-CTLA4Ig chimeric molecule and its expression on CHO-dhfr(-) cells].
abstract: OBJECTIVE
  To develop a novel immunosuppressant GPI-CTLAIg modified by glycosyl-phosphatidylinositol(GPI).
  
  
  METHODS
  GPI-modified CTLAIg was produced by linking-up of CTLA4Ig with GPI-modification signal sequences from decay-accelerating factor (DAF). Chimeric molecule GPI-CTLA4Ig gene was cloned into eukaryotic expression vector pCI-dhfr. Using lipofectine-mediated gene transfer technique, pCI-GPI-CTLA4Ig was transfected into CHO-dhfr(-) cells, and the transfectants were screened by methotrexate (MTX). Expression of the recombinant protein was assessed by RT-PCR, ELISA, cell immunofluorescence staining and Western blot, and the purification of expressed protein was performed by protein A affinity chromatography.
  
  
  RESULTS
  The chimeric molecule GPI-CTLA4Ig has been constructed and stablely expressed on CHO-dhfr(-) cells.
  
  
  CONCLUSIONS
  GPI-modified CTLAIg will may be used as novel immunosuppressant for suppressing reaction in graft rejection.
authors.affiliation: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850,China.
authors.forename: Ji-Yun
authors.initials: JY
authors.lastname: Yu
authors.forename: Bei-Fen
authors.initials: BF
authors.lastname: Shen
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Xing
authors.initials: X
authors.lastname: Chen
authors.forename: Shao-Hui
authors.initials: SH
authors.lastname: Cheng
authors.forename: Rong
authors.initials: R
authors.lastname: Tian
authors.forename: Mei-Hua
authors.initials: MH
authors.lastname: Qu
mesh_terms: D000818:Animals; D018958:CD55 Antigens; D016466:CHO Cells; D018014:Gene Transfer Techniques; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
chemical_list: D018958:CD55 Antigens; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins