Cook, A., Hari-Gupta, Y. and Toseland, C.P. orcid.org/0000-0002-1641-7535 (2018) Application of the SSB biosensor to study in vitro transcription. Biochemical and Biophysical Research Communications, 496 (3). pp. 820-825. ISSN 0006-291X
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.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | © 2018 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | Biosensor; Transcription; SSB; Myosin; RNA polymerase; Gene expression; mRNA |
Dates: |
|
Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Oncology (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 13 Nov 2019 14:19 |
Last Modified: | 13 Nov 2019 14:19 |
Status: | Published |
Publisher: | Elsevier BV |
Refereed: | Yes |
Identification Number: | 10.1016/j.bbrc.2018.01.147 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:152398 |