The novel molecule porcine OSR1 up-regulated expression on porcine endothelial cell by human peripheral blood mononuclear cell activation.
BACKGROUND
Clinical xenotransplantation is being considered to overcome the shortage of human organ donors. In our previous studies, using human-anti-porcine xenogeneic mixed mononuclear cell-endothelial cell cultures with the suppression subtractive hybridization method, we obtained a subtracted cDNA library that included about 300 clones corresponding to up-regulated genes. One porcine sequence showed 81% identity with the human oxidative-stress responsive 1 (OSR1) molecule. The objective of this study was to confirm the gene up-regulation and obtain the full-length sequences.


METHODS
The full-length gene was cloned through the technique of rapid amplification of cDNA ends (RACE). The other methods included bioinformatics analysis and RT-PCR.


RESULTS
RT-PCR confirmed that the gene was up-regulated upon the interactions of human peripheral blood mononuclear cells (PBMCs) and porcine endothelial cells. By SMART RACE technique, we obtained the full-length cDNA of porcine OSR1. The gene is 4333 bp. The open-read frame of 1590 bp encodes 529 amino acid residues. GenBank accession number is AY271356. The gene shows 92.8% nucleotide identity and 95.5% amino acid identity with human OSR1.


CONCLUSIONS
We obtained the full-length cDNA of porcine OSR1. It was up-regulated on porcine endothelial cells following activation by human PBMCs. We succeeded in constructing a pcDNA-pOSR1 recombinant eukaryotic cell expression vector, the function of which is the subject as our ongoing work.
Hu
W
W
Lab of Transplant Engineering and Immunology, West China Hospital, Sichuan University, People's Republic of China.
Cheng
J
J
Lu
X
X
Li
S
S
Zeng
L
L
Li
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004730:Endothelium, Vascular / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000502:physiology*; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013552:Swine
C118135:OXSR1 protein, human; D017346:Protein Serine-Threonine Kinases