In vitro experimental system for analysis of transcription-translation coupling


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.


PubMed ID: 22210860

Projects: Noisy-Strep

Publication type: Not specified

Journal: Nucleic acids research


Date Published: 3rd Jan 2012

Registered Mode: Not specified

Authors: Daniel Castro-Roa,

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Views: 3685

Created: 5th Jan 2012 at 18:00

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