Eukaryotic expression vector of heme oxygenase-1 and its expression in endothelial cell

BACKGROUND: Heme oxygenase-1 (HO-1)-mediated cy-toprotection may inhibit or postpone the formation of graftarteriosclerosis. The aim of this study was to construct eu-karyotic expression vector of HO-1 gene and express re-combinant HO-1 in human endothelial cells of the umbili-cal vein to provide an important clue for chronic rejectionstudy.METHODS: HO-1 gene was amplified from rat spleen withreverse transcription-polymerase chain reaction, and thencloned into eukaryotic expression vector pcDNA3 by meansof recombinant gene technology. The recombinant plasmidpcDNA3-HO-l was transfected into endothelial cells, andrecombinant HO-1 was expressed in the endothelial cellsunder G418 selection. The expressed products were detec-ted using indirect fluorescent staining and Western blotanalysis.RESULTS: Restriction analysis indicated that the HO-1gene was successfully inserted into the expression vector,and DNA sequencing verified that the reading frame of therecombinant vector was correct. Recombinant HO-1 wassuccessfully expressed in endothelial cells and its molecularweight was about 32 kD.CONCLUSION: The successful construction of eukaryoticexpression vector containing the HO-1 gene and the effec-tive expression of recombinant HO-1 in endothelial cellshas laid a foundation for further study on its biologicalfunctions.