Chloroform-free permeabilization for improved detection of β-galactosidase activity in Vibrio cholerae

LacZ (β-galactosidase) is used to monitor the transcription of genes in reporter strains carrying the lacZ gene under the control of a promotor of interest. This protocol for LacZ activity determinations in Vibrio cholerae following detergent lysis results in 2.5-fold increase of LacZ activities com...

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Bibliographic Details
Published in:Journal of microbiological methods 2017-06, Vol.137, p.1-2
Main Authors: Toulouse, Charlotte, Häse, Claudia C., Steuber, Julia
Format: Article
Language:English
Subjects:
Bacterial cell lysis
Bacterial Proteins - genetics
beta-Galactosidase - genetics
beta-Galactosidase - metabolism
Cell Membrane - metabolism
Chloroform - pharmacology
Detergents - pharmacology
Genes, Reporter
Lac Operon
LacZ
Nitrophenols - metabolism
Promoter Regions, Genetic
Protease Inhibitors - pharmacology
Transcription, Genetic
Vibrio cholerae
Vibrio cholerae - drug effects
Vibrio cholerae - enzymology
Vibrio cholerae - genetics
β-Galactosidase
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Summary:LacZ (β-galactosidase) is used to monitor the transcription of genes in reporter strains carrying the lacZ gene under the control of a promotor of interest. This protocol for LacZ activity determinations in Vibrio cholerae following detergent lysis results in 2.5-fold increase of LacZ activities compared to lysis with chloroform. •Detergent lysis buffer for disruption of bacterial cells in 96-well format•Optimization of protein:detergent ratio for cell lysis•Increased β-galactosidase activity compared to lysis in chloroform
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2017.03.011