ox-LDL特异性人血管平滑肌细胞cDNA消减文库的构建

Establishment of Subtractive cDNA library specific to ox - LDL

目的:构建ox-LDL特异性人血管平滑肌细胞cDNA消减文库。方法:35％ug/ml ox-LDL刺激培养的人血管平滑肌细胞,利用消减杂交技术,构建cDNA消减文库,并运用蓝白斑筛选和影印杂交进一步确定文库克隆的特异性。结果:成功构建ox-LDL特异性血管平滑肌细胞cDNA消减文库,约2000个白色克隆。影印杂交后共获82个差异表达克隆。结论:消减文库的构建为进一步克隆ox-LDL特异性基因cDNA片段和全长奠定了基础。

objective to establish the subtractive cDNA library specific to ox - LDL. Methods ox - LDL was produced and added as a stimulant to SMC culture medium , using Subtractive hybridization technique, bluewhite clone screening system and blotting hybridization , we set up the subtractive cDNA library. Results The white clones amount up to 2000, differentiati library exlpressed clone were about 82. Conclusion The Subtractive Library was the foundation for cloning the whole - length c DNA and Its function study.