高通量筛选DNA加合物位点方法的探讨

Study on a high throughput method of screening adducted site of DNA adducts

目的探讨建立一种高通量的筛选DNA加合物结合位点的方法。方法以烷化剂氮芥处理人肝瘤HepG2细胞形成DNA加合物模型,利用基因表达系列分析技术(serrialanalysisofgeneexpression,SAGE)对连接介导聚合酶链式反应(ligationmediatedPCR,LMPCP)进行改良,以改良后的方法进行检测。结果PCR扩增产物大小约为75bp,与理论设计相符合。结论该方法的设计合理、可行。

Objective To develop a high throughput method of screening the binding site of DNA adducts.Methods The human liver tumor HepG2 cells were first treated with lng/μl nitrogen mustard for 3h and total DNA was extracted,then serrial analysis of gene expression was used for reference to improve ligation mediated PCR to detect the adducted site by nitrogen mustard in the cell genome.Results PCR product corresponded to 75 base pairs approximately.Conclusion The design of the experiment is rational and feasible.