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.
SEEK ID: https://fairdomhub.org/publications/132
PubMed ID: 22210860
Projects: Noisy-Strep
Publication type: Not specified
Journal: Nucleic acids research
Citation:
Date Published: 3rd Jan 2012
Registered Mode: Not specified
Views: 5420
Created: 5th Jan 2012 at 17:00
Last updated: 8th Dec 2022 at 17:26
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