Screening of atherosclerosis related genes using suppression subtractive hybri dization technique

AIM To screen the atherosclerosis related genes. METH OD S The differentially expressed subtracted cDNA library of freshly isolate d adult bovine endothelium and cultured new-born bovine endothelium constructed by suppression subtractive hybridization technique was screened by colony in situ hybridization, the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the GenBank databases with basic local alignment search tool. The atherosclerosi s related genes were analyzed by RNA dot-blot analysis on rabbit atherosclerosi s model. RESULTS Eighty-eight positive clones were obtained, and 6 1 cDNA sequences were identified. Sequences of 61 cDNA showed that 26 cDNA were known as bovine gene sequences, 19 cDNA were homologous with the human g enes published in GenBank and 16 cDNA were unknown genes. Through gene homologou s analysis, some differentially expressed cDNA sequences were selected for tissu e differential and atherosclerosis related analysis. We found that DEAD-box pro tein, factor Ⅻa inhibitor, lymphocyte adaptor protein and protein translocation complex β are all atherosclerosis related genes. CONCLUSION This is the first time to report four genes are related to atherosclerosis.