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

Analytical Biochemistry
Enrico MonachinoAntoine M van Oijen

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.

Citations

Feb 16, 2019·Nucleic Acids Research·Lisanne M SpenkelinkAntoine M van Oijen
May 7, 2020·Nucleic Acids Research·Katarzyna DubielJames L Keck
Sep 19, 2019·Scientific Reports·Kelsey S WhinnHarshad Ghodke
Nov 26, 2020·Essays in Biochemistry·Gurleen Kaur, Lisanne M Spenkelink
May 29, 2020·Molecular Cell·Enrico MonachinoAntoine M van Oijen
Jun 18, 2021·Nucleic Acids Research·Richard R SpinksAntoine M van Oijen

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