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A+G连接介导PCR方法的建立及其在体内足迹研究中的应用
Establishment of A+G Ligation Mediated Polymerase Chain Reaction and Its Application in in vivo Footprinting Study
【摘要】 目的建立一种新的连接介导聚合酶链反应(ligation-mediatedpoly-merasechainreaction,LM-PCR)方法,使之能够对G-残基含量过低或不含G-残基的区域进行体内足迹研究。方法对原化学测序法中的A>G法进行修改,化学裂解后进行连接介导的PCR反应,通过测序胶分离分析PCR反应结果。结果建立了A+GLM-PCR方法,本法能够应用于体内足迹研究。结论应用本实验建立的方法可分析A-残基和G-残基上的蛋白质-DNA相互作用状况,可获得更多信息及扩大使用范围。
【Abstract】 Objective To establish a new ligation mediated PCR method for the work in in vivo footprinting study of some regions of low G residue content or some regions of G residue free. Methods The A>G chemical cleavage method of Maxam Gilbert sequencing was modified for ligation mediated PCR after A+G chemical cleavage and separate the PCR products by sequencing PAGE. Results A new A+G ligation mediated method was established and could be used in in vivo footprinting study. Conclusions Our results indicated that the newly developed method could analyze the DNA Protein interaction at both A residue and G residue,enabled us to get more information and enlarged the scope of its application.
【Key words】 ligation mediated PCR; A>G chemical cleavage; sequencing; dimethyl sulfate; in vivo footprinting;
- 【文献出处】 中国医学科学院学报 ,ACTA ACADEMIAE MEDICINAE SINICAE , 编辑部邮箱 ,1999年03期
- 【分类号】Q5
- 【下载频次】115