内皮_血管_细胞学；抑制消减杂交；动脉硬化_

传代培养前后内皮细胞差异表达基因消减cDNA文库的构建

http://www.100md.com 《第四军医大学学报》 2006年第12期

内皮,血管,细胞学；抑制消减杂交；动脉硬化,,内皮，血管,细胞学；抑制消减杂交；动脉硬化,0引言,1材料和方法,2结果,3讨论,【参考文献

     【Abstract】 AIM: To construct a subtracted cDNA library of differentially expressed genes in freshly isolated adult bovine aortic endothelial cells (FiBEC) and cultured newborn aortic endothelial cells(CnBEC). METHODS: The suppression subtractive hybridization (SSH) was used to isolate cDNA fragments of nonexpressed or lowexpressed 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 arteriosclerosisrelated genes in early stage.

    【Keywords】 endothelium; vascular/cytology; suppression subtractive hybridization; arteriosclerosis

    【摘要】目的：构建传代培养前后牛主动脉内皮细胞差异表达基因的消减cDNA文库，筛选动脉粥样硬化相关新基因构建文库. 方法：采用抑制消减杂交技术，以新鲜分离和传代培养的牛主动脉内皮细胞作为对比材料，分离传代培养主动脉内皮细胞中不表达或低表达基因的cDNA片段 ......

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