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Methods for the Assembly and Analysis of In Vitro Transcription-Coupled-to-Translation Systems

Lookup NU author(s): Dr Daniel Castro Roa, Professor Nikolay ZenkinORCiD


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RNA polymerase is a complex machinery, which is further embedded in interactions with other cellular components that interplay with either the transcribed DNA (DNA polymerases, topoisomerases, etc.) or the nascent RNA (RNA processing enzymes, ribosomes, etc.). In prokaryotes, coupling of transcription and translation is thought to play many regulatory roles but the mechanistic understanding of their interactions has been hindered by the lack of a defined experimental system. Here, we describe a pure transcription-coupled-to-translation system in which control of the ribosome has been achieved through its stepwise translocation towards RNA polymerase. This system can be used to study the effects of concurrent translation on RNA chain elongation and to elucidate the interface between the two macromolecular complexes.

Publication metadata

Author(s): Castro-Roa D, Zenkin N

Editor(s): Irina Artsimovitch and Thomas J. Santangelo

Publication type: Book Chapter

Publication status: Published

Book Title: Bacterial Transcriptional Control

Year: 2015

Volume: 1276

Pages: 81-99

Print publication date: 28/01/2015

Online publication date: 21/02/2015

Series Title: Methods in Molecular Biology

Publisher: Springer New York

Place Published: New York, NY. USA


DOI: 10.1007/978-1-4939-2392-2_5

PubMed id: 25665559

Library holdings: Search Newcastle University Library for this item

ISBN: 9781493923915