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 ,经T载体转化感受态大肠杆菌。结果 :建立了内皮细胞内毒素刺激后差异表达基因消减cDNA文库。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 。
Objective:To construct the subtractive differentially expressed genes libraries in human umbilical vein endothelial cells (HUVECs) treated by endotoxin.Methods:After culturing HUVECs treated or not with endotoxin for 6 hours, their mRNAs were isolated to synthesize cDNAs by reverse transcription. The differentially expressed cDNAs were enriched by suppression subtractive hybridization (SSH), directly inserted a T vector and transformed into DH5a E.Coli competent cells.Results:Constructing subtractive cDNA libraries in HUVECs.Conclusion:The libraries from SSH enriched and equalized the differentially expressed genes varied in abundance, which made it convenient to screen LPS-associatied genes.
出处
《西北国防医学杂志》
CAS
2002年第2期81-84,共4页
Medical Journal of National Defending Forces in Northwest China
基金
国家自然科学基金重点资助项目 (3 983 0 40 0 )
