固有弯曲DNA的体外进化

In vitro evolution of intrinsically bent DNA.

作者信息

Beutel B A, Gold L

机构信息

Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309-0347.

出版信息

J Mol Biol. 1992 Dec 5;228(3):803-12. doi: 10.1016/0022-2836(92)90865-h.

DOI:10.1016/0022-2836(92)90865-h

PMID:1469714

Abstract

DNA fragments which are intrinsically bent or curved migrate anomalously during electrophoresis through polyacrylamide gels. Starting with an initial population of approximately 10(12) unique DNA sequences, DNA which exhibited the kind of anomalous mobility associated with DNA bending was selected and enriched using a variation of the SELEX procedure. After seven rounds of selection and amplification, the vast majority of the remaining population of DNA fragments migrated as bent DNA. Cloning and sequencing of 30 individual sequences from this population has yielded information regarding the relationship between DNA sequence and bending. Some of the previous conclusions on DNA bending have been confirmed while others have been modified, by the results presented here. In addition, the dinucleotide base step CA/TG, which had not been thought to be a major factor in DNA bending, appears to be important.

摘要

本质上发生弯曲或卷曲的DNA片段在通过聚丙烯酰胺凝胶进行电泳时会出现异常迁移。从大约10¹²个独特DNA序列的初始群体开始，选择表现出与DNA弯曲相关的那种异常迁移率的DNA，并使用SELEX程序的一种变体进行富集。经过七轮选择和扩增后，剩余的绝大多数DNA片段群体迁移时表现为弯曲的DNA。对该群体中30个单独序列进行克隆和测序，得出了有关DNA序列与弯曲之间关系的信息。此处给出的结果证实了先前关于DNA弯曲的一些结论，同时也对其他一些结论进行了修正。此外，以前未被认为是DNA弯曲主要因素的二核苷酸碱基步CA/TG似乎很重要。