设计具有可控叉状拓扑结构的DNA滚环模板以研究DNA复制机制。

Design of DNA rolling-circle templates with controlled fork topology to study mechanisms of DNA replication.

作者信息

Monachino Enrico, Ghodke Harshad, Spinks Richard R, Hoatson Ben S, Jergic Slobodan, Xu Zhi-Qiang, Dixon Nicholas E, van Oijen Antoine M

机构信息

Molecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, and Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia; Zernike Institute for Advanced Materials, University of Groningen, 9747 AG, Groningen, The Netherlands.

Molecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, and Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia.

出版信息

Anal Biochem. 2018 Sep 15;557:42-45. doi: 10.1016/j.ab.2018.07.008. Epub 2018 Jul 18.

DOI:10.1016/j.ab.2018.07.008

PMID:30016625

Abstract

Rolling-circle DNA amplification is a powerful tool employed in biotechnology to produce large from small amounts of DNA. This mode of DNA replication proceeds via a DNA topology that resembles a replication fork, thus also providing experimental access to the molecular mechanisms of DNA replication. However, conventional templates do not allow controlled access to multiple fork topologies, which is an important factor in mechanistic studies. Here we present the design and production of a rolling-circle substrate with a tunable length of both the gap and the overhang, and we show its application to the bacterial DNA-replication reaction.

摘要

滚环式DNA扩增是生物技术中用于从少量DNA大量生产DNA的一种强大工具。这种DNA复制模式通过类似于复制叉的DNA拓扑结构进行，因此也为研究DNA复制的分子机制提供了实验途径。然而，传统模板不允许对多种叉状拓扑结构进行可控访问，而这是机制研究中的一个重要因素。在此，我们展示了一种间隙和突出端长度均可调的滚环底物的设计与生产，并展示了其在细菌DNA复制反应中的应用。