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In vitro experimental system for analysis of transcription-translation coupling

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

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Abstract

Transcription and translation are coupled in bacteria, meaning that translation takes place co-transcriptionally. During transcription–translation, both machineries mutually affect each others’ functions, which is important for regulation of gene expression. Analysis of interactions between RNA polymerase (RNAP) and the ribosome, however, are limited due to the lack of an in vitro experimental system. Here, we report the development of an in vitro transcription coupled to translation system assembled from purified components. The system allows controlled stepwise transcription and simultaneous stepwise translation of the nascent RNA, and permits investigation of the interactions of RNAP with the ribosome, as well as the effects of translation on transcription and transcription on translation. As an example of usage of this experimental system, we uncover complex effects of transcription–translation coupling on pausing of transcription.

Publication metadata

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

Publication type: Article

Publication status: Published

Journal: Nucleic Acids Research

Year: 2012

Volume: 40

Issue: 6

Print publication date: 30/12/2011

Date deposited: 12/10/2012

ISSN (print): 0305-1048

ISSN (electronic): 1362-4962

Publisher: Oxford University Press

URL: http://dx.doi.org/10.1093/nar/gkr1262

DOI: 10.1093/nar/gkr1262

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