摘要
目的:建立人脐静脉内皮细胞内毒素刺激后差异表达基因的消减cDNA文库。方法:提取培养人脐静脉内皮细胞内毒素刺激6h后mRNAs,反转录合成cDNA,与对照组间进行抑制消减杂交富集差异表达基因,亚克隆入T/A质粒并转化感受态大肠杆菌。结果:成功构建了人脐静脉内皮细胞内毒素刺激后上调和下调差异表达基因两个消减cDNA文库,为进一步筛选新的内皮细胞活化相关基因打下基础。结论:经抑制消减杂交建立的消减cDNA文库富集并均一化了内毒素刺激后不同丰度的差异表达基因,对深入理解内皮细胞活化机制及参与组织修复的过程具有重要意义。
AIM:To construct a subducting cDNA library of differentially expressed genes after the stimulation of endotoxin to human umbilical vein endothelial cells(HUVECs). METHODS:mRNAs of cultured HUVECs stimulated by endotoxin after 6h were extracted, and then cDNA was synthesized by reverse transcription. Suppressive subducting hybridization was used to enrich the differential expression of genes.T/A plasmid was subcloned and competence escherichia coli was conversed. RESULTS:Two subducting cDNA libraries of up regulation and down regulation differentially expressed genes in HUVECs after stimulated by endotoxin were constructed successfully.It was the basis for screening new endothelial cells to activate the related genes. CONCLUSION:The subducting cDNA library constructed by suppressive subducting hybridization enriched and homogenized the differentially expressed genes of different degrees of abundance stimulated by endotoxin. It is important for understanding the activation mechanism of endothelial cells and their role in the process of tissue repair.
出处
《中国临床康复》
CSCD
2004年第5期972-973,T004,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然基金青年基金(30200101)~~
