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Single-molecule observation of prokaryotic DNA replication

Journal Article


Abstract


  • Recent advances in optical imaging and molecular manipulation techniques have made it possible to observe the activity of individual enzymes and study the dynamic properties of processes that are challenging to elucidate using ensemble-averaging techniques. The use of single-molecule approaches has proven to be particularly successful in the study of the dynamic interactions between the components at the replication fork. In this section, we describe the methods necessary for in vitro single-molecule studies of prokaryotic replication systems. Through these experiments, accurate information can be obtained on the rates and processivities of DNA unwinding and polymerization. The ability to monitor in real time the progress of a single replication fork allows for the detection of short-lived, intermediate states that would be difficult to visualize in bulk-phase assays.

Publication Date


  • 2009

Citation


  • Tanner, N. A. & van Oijen, A. M. (2009). Single-molecule observation of prokaryotic DNA replication. Methods in Molecular Biology, 521 397-410.

Scopus Eid


  • 2-s2.0-68349146847

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2204

Has Global Citation Frequency


Number Of Pages


  • 13

Start Page


  • 397

End Page


  • 410

Volume


  • 521

Place Of Publication


  • United States

Abstract


  • Recent advances in optical imaging and molecular manipulation techniques have made it possible to observe the activity of individual enzymes and study the dynamic properties of processes that are challenging to elucidate using ensemble-averaging techniques. The use of single-molecule approaches has proven to be particularly successful in the study of the dynamic interactions between the components at the replication fork. In this section, we describe the methods necessary for in vitro single-molecule studies of prokaryotic replication systems. Through these experiments, accurate information can be obtained on the rates and processivities of DNA unwinding and polymerization. The ability to monitor in real time the progress of a single replication fork allows for the detection of short-lived, intermediate states that would be difficult to visualize in bulk-phase assays.

Publication Date


  • 2009

Citation


  • Tanner, N. A. & van Oijen, A. M. (2009). Single-molecule observation of prokaryotic DNA replication. Methods in Molecular Biology, 521 397-410.

Scopus Eid


  • 2-s2.0-68349146847

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2204

Has Global Citation Frequency


Number Of Pages


  • 13

Start Page


  • 397

End Page


  • 410

Volume


  • 521

Place Of Publication


  • United States