title: [Construction of recombinant human CD59 using ICAM-2 promoter for endothelial-specific expression in xenotransplantation].
abstract: OBJECTIVE
  To construct a recombinant human CD59 gene containing intercellular adhesion molecule-2 promoter for high level endothelial-specific expression in xenotransplantation.
  
  
  METHODS
  ICAM-2 promotor fragment and CD59-intron 1 fragment were produced by PCR from the human blood genome, and then clone these fragments into a pcDNA3-CD59 eukaryotic expression vector which was followed by digestion with the specific restricted endonuclease (for example: EcoRI, Hind III). The ICAM-2 promoter and CD59-intron 1 fragments were identified by PCR, and sequencing. The recombinant was then transfected into pig aorta endothelial cells with Lipofection, and the expression was measured by flow cytometer.
  
  
  RESULTS
  Products of the sequences measured were in accord with the frames of the gene bank. The expression of the protein of this recombinant was positive.
  
  
  CONCLUSIONS
  The CD59 recombinant gene is constructed successfully, providing a basis for transgenic research.
authors.affiliation: Tianjin Institute of Urology Surgery, Tianjin Medical University, Tianjin 300211, China. yaoxu7962_cn@sina.com
authors.forename: Xu-dong
authors.initials: XD
authors.lastname: Yao
authors.forename: Teng-xiang
authors.initials: TX
authors.lastname: Ma
authors.forename: Zhi-hong
authors.initials: ZH
authors.lastname: Wu
authors.forename: Sheng-zhi
authors.initials: SZ
authors.lastname: Li
authors.forename: Zhi-xin
authors.initials: ZX
authors.lastname: Li
authors.forename: Guang-you
authors.initials: GY
authors.lastname: Wang
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D018957:CD59 Antigens / Q000096:biosynthesis / Q000235:genetics*; D015815:Cell Adhesion Molecules / Q000235:genetics*; D003001:Cloning, Molecular; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013552:Swine; D014162:Transfection; D014183:Transplantation, Heterologous
chemical_list: D015703:Antigens, CD; D018957:CD59 Antigens; D015815:Cell Adhesion Molecules; C065289:ICAM2 protein, human; D011994:Recombinant Proteins