Transplantation proceedings · 2004
The novel molecule porcine OSR1 up-regulated expression on porcine endothelial cell by human peripheral blood mononuclear cell activation.
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Hu W, Cheng J, Lu X, Li S, Zeng L, Li Y
Аннотация
INTRODUCTION: 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.
CONCLUSION: 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.
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.
CONCLUSION: 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.