基于报告基因能筛选下调p66shc基因转录的药物普筛系统的建立
Establishment of a Reporter Gene-Based Drug Screening System to Identify Down-Regulators to the Transcriptional Regulation of p66shc Gene
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摘要: 将p66shc基因的启动子序列克隆到含有荧光素酶报告基因和新霉素抗性基因的pGL3B-neo载体中,然后将重组质粒pGL3B-neo-p66shc转染入人的肺癌细胞A549中,通过单集落分离法得到含有重组质粒的稳定细胞系A549/p66shc。利用这个以报告基因为基础的药物普筛系统,筛选了数百种中药提取物及单体化合物,发现抑制率达到50%以上的2种中药提取物和8种单体化合物。Abstract: The promoter of p66~(shc) gene was cloned into the pGL3B-neo vector containing the luciferase gene and neomycin resistance gene,and the recombinant vector pGL3B-neo-p66~(shc) was subsequently transfected into A549 cells,and then the stable cell line A549/p66~(shc) was obtained through single colony iso-(lation.) Using this reporter gene-based screening system,several hundred samples were screened and ten of them were found to meet demands.