| 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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| 引用本文： | 张萍,山丽梅,肖小河,付强.传代培养前后内皮细胞差异表达基因消减cDNA文库的构建[J].第四军医大学学报,2006,27(12):1100-1103. |
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| 作者姓名： | 张萍 山丽梅 肖小河 付强 |
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| 作者单位： | 1. 军事医学科学院毒物药物研究所,北京,100850
2. 解放军第302医院中医药研究所,北京,100039 |
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| 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,将其与T载体进行T/A连接构建文库,将连接产物转化大肠杆菌进行文库扩增后,随机挑取64个白色克隆进行酶切鉴定. 结果:扩增消减cDNA文库获得760个克隆,随机挑取的64个阳性克隆经酶切后均有200～600 bp的插入片段. 结论:构建的传代培养前后内皮细胞差异表达基因消减cDNA文库,为进一步筛选早期发生改变的动脉粥样硬化相关基因奠定了工作基础.
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| 关 键 词： | 内皮 血管/细胞学 抑制消减杂交 动脉硬化 |
| 文章编号： | 1000-2790(2006)12-1100-04 |
| 收稿时间： | 02 28 2006 12:00AM |
| 修稿时间： | 04 3 2006 12:00AM |
Construction of a subtracted cDNA library of differentially expressed genes in freshly isolated adult and cultured new-born bovine aortic endothelial cells |
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| ZHANG Ping,SHAN Li-Mei,XIAO Xiao-He,FU Qiang.Construction of a subtracted cDNA library of differentially expressed genes in freshly isolated adult and cultured new-born bovine aortic endothelial cells[J].Journal of the Fourth Military Medical University,2006,27(12):1100-1103. |
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| Authors: | ZHANG Ping SHAN Li-Mei XIAO Xiao-He FU Qiang |
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| Affiliation: | 1 Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China, 2 Institute of Traditional Chinese Medicine, 302 Hospital of PLA, Beijing 100039, China |
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| 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. |
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| Keywords: | endothelium vascular/cytology suppression subtractive hybridization arteriosclerosis |
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