Colony Lift Colorimetric Assay for beta-Galactosidase Activity
Program in Systems Biology; Program in Molecular Medicine; UMass Metabolic Network
Genetics and Genomics | Laboratory and Basic Science Research | Molecular Biology | Systems Biology
In this protocol, we present a qualitative assay for monitoring the level of expression of beta-galactosidase, an enzyme encoded by the LacZ gene, in yeast. This is useful both for determining autoactivity of LacZ expression in yeast DNA "bait" strains and for assessing LacZ reporter gene activation mediated by a transcription factor "prey" interaction with a DNA bait of interest in yeast one-hybrid (Y1H) assays. In this colorimetric assay, yeast are lysed in liquid nitrogen and then assayed for beta-galactosidase expression using the colorless compound X-gal, which turns blue in the presence of this enzyme.
DOI of Published Version
Cold Spring Harb Protoc. 2016 Dec 1;2016(12):pdb.prot088963. doi: 10.1101/pdb.prot088963 Link to article on publisher's site
Cold Spring Harbor protocols
Fuxman Bass, Juan; Reece-Hoyes, John S.; and Walhout, Albertha J. M., "Colony Lift Colorimetric Assay for beta-Galactosidase Activity" (2016). Program in Systems Biology Publications and Presentations. 105.