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毛细管电泳在配体 - DNA相互作用研究中的应用。

Use of capillary electrophoresis in the study of ligand-DNA interactions.

作者信息

Hamdan I I, Skellern G G, Waigh R D

机构信息

Department of Pharmaceutical Sciences, University of Strathclyde, Glasgow G1 1XW, UK.

出版信息

Nucleic Acids Res. 1998 Jun 15;26(12):3053-8. doi: 10.1093/nar/26.12.3053.

DOI:10.1093/nar/26.12.3053
PMID:9611254
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC147652/
Abstract

Free solution capillary electrophoresis (FSCE) has been used to separate two non-self-complementary 12mer oligonucleotide duplexes: d(AAATTATATTAT).d(ATAA-TATAATTT) and d(GGGCCGCGCCGC).d(GCGGCGCGGCCC). Titration of mixtures of the two oligonucleotides with model intercalators (ethidium bromide andactinomycin D) and minor groove binders (netropsin, Hoechst 33258 and distamycin) has shown the suitability of FSCE as a method to study the sequence selectivity of DNA binding agents. Binding data have shown cooperativity of binding for netropsin and Hoechst 33258 and have provided ligand:DNA binding ratios for all five compounds. Cooperativity of netropsin binding to a 12mer with two potential sites has been demonstrated for the first time. Ligands binding in the minor groove caused changes in migration time and peak shape which were significantly different from those caused by intercalators.

摘要

自由溶液毛细管电泳（FSCE）已被用于分离两种非自互补的12聚体寡核苷酸双链体：d(AAATTATATTAT).d(ATAA-TATAATTT) 和 d(GGGCCGCGCCGC).d(GCGGCGCGGCCC)。用模型嵌入剂（溴化乙锭和放线菌素D）和小沟结合剂（纺锤菌素、Hoechst 33258和偏端霉素）对这两种寡核苷酸的混合物进行滴定，结果表明FSCE适合作为研究DNA结合剂序列选择性的方法。结合数据显示了纺锤菌素和Hoechst 33258结合的协同性，并提供了所有五种化合物的配体与DNA的结合比率。首次证明了纺锤菌素与具有两个潜在位点的12聚体结合的协同性。在小沟中结合的配体导致迁移时间和峰形的变化，这些变化与嵌入剂引起的变化显著不同。

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