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Application of the SSB biosensor to study in vitro transcription

Cook, Alexander, Hari-Gupta, Yukti, Toseland, Christopher P. (2018) Application of the SSB biosensor to study in vitro transcription. Biochemical and Biophysical Research Communications, 496 (13). pp. 820-825. ISSN 0006-291X. (doi:10.1016/j.bbrc.2018.01.147)

Abstract

Gene expression, catalysed by RNA polymerases (RNAP), is one of the most fundamental processes in living cells. The majority of methods to quantify mRNA are based upon purification of the nucleic acid which leads to experimental inaccuracies and loss of product, or use of high cost dyes and sensitive spectrophotometers. Here, we describe the use of a fluorescent biosensor based upon the single stranded binding (SSB) protein. In this study, the SSB biosensor showed similar binding properties to mRNA, to that of its native substrate, single-stranded DNA (ssDNA). We found the biosensor to be reproducible with no associated loss of product through purification, or the requirement for expensive dyes. Therefore, we propose that the SSB biosensor is a useful tool for comparative measurement of mRNA yield following in vitro transcription.

Item Type: Article
DOI/Identification number: 10.1016/j.bbrc.2018.01.147
Uncontrolled keywords: Biosensor; Transcription; SSB; Myosin; RNA polymerase; Gene expression; mRNA
Subjects: Q Science
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Chris Toseland
Date Deposited: 02 Feb 2018 16:18 UTC
Last Modified: 29 May 2019 20:13 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/65865 (The current URI for this page, for reference purposes)
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