Assays for transcription factor activity

Douglas Browning; Nigel Savery; Annie Kolb; Stephen Busby

doi:10.1007/978-1-60327-015-1_23

Assays for transcription factor activity

Methods Mol Biol. 2009:543:369-87. doi: 10.1007/978-1-60327-015-1_23.

Authors

Douglas Browning¹, Nigel Savery, Annie Kolb, Stephen Busby

Affiliation

¹ School of Biosciences, The University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.

PMID: 19378177
DOI: 10.1007/978-1-60327-015-1_23

Abstract

Transcription factors interact at promoters to modulate the transcription of genes. This chapter describes three in vitro methods that can be used to monitor their activity: transcript assays, abortive initiation assays, and potassium permanganate footprinting. These techniques have been developed using bacterial systems, and can be used to study the kinetics of transcription initiation, and hence to unravel regulatory mechanisms.

MeSH terms

Biological Assay / methods*
Cyclic AMP Receptor Protein / metabolism
DNA Footprinting
DNA-Directed RNA Polymerases / metabolism
Escherichia coli / enzymology
Escherichia coli / genetics
Plasmids / genetics
Potassium Permanganate
Promoter Regions, Genetic / genetics
RNA, Messenger / genetics
RNA, Messenger / metabolism
Transcription Factors / metabolism*
Transcription, Genetic
Transcriptional Activation / genetics

Substances

Cyclic AMP Receptor Protein
RNA, Messenger
Transcription Factors
Potassium Permanganate
DNA-Directed RNA Polymerases