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传代培养前后内皮细胞差异表达基因消减cDNA文库的构建
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【西医学论文】 作者:张萍,山丽梅,肖小河,付强【关键词】 内皮,血管/细胞学;抑制消减杂交;动脉硬化【Abstract】 AIM: To construct a subtracted cDNA library of differentially expressed genes in freshly isolated adult bovine aortic endothelial cells (FiBEC) and cultured newborn aortic endothelial cells(CnBEC). METHODS: The suppression subtractive hybridization (SSH) was used to isolate cDNA fragments of nonexpressed or lowexpressed genes in CnBEC . Then these cDNA fragments were directly inserted into T/A cloning vestor to set up the subtractive library. The amplified library was randomly picked and identified using restriction enzyme digestion. RESULTS: The amplified library contained more than 760 bacterial clones. The 64 clones picked randomly showed that all clones contained 200-600 bp inserts after restriction enzyme digestion. CONCLUSION: The library lays a foundation for screening and cloning the
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