[Construction of recombinant human CD59 using ICAM-2 promoter for endothelial-specific expression in xenotransplantation].
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.
Yao
Xu-dong
XD
Tianjin Institute of Urology Surgery, Tianjin Medical University, Tianjin 300211, China. yaoxu7962_cn@sina.com
Ma
Teng-xiang
TX
Wu
Zhi-hong
ZH
Li
Sheng-zhi
SZ
Li
Zhi-xin
ZX
Wang
Guang-you
GY
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
D015703:Antigens, CD; D018957:CD59 Antigens; D015815:Cell Adhesion Molecules; C065289:ICAM2 protein, human; D011994:Recombinant Proteins