Fluorescence-based detection oflacZreporter gene expression in intact and viable bacteria includingMycobacteriumspecies
Open Access
- 1 October 1999
- journal article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 179 (2), 317-325
- https://doi.org/10.1111/j.1574-6968.1999.tb08744.x
Abstract
A variety of fluorescein di-β-d-galactopyranoside (FDG)-based substrates were evaluated for measuring β-galactosidase expression in bacteria. One substrate, 5-acetylamino-FDG (C2FDG), performed well in all bacteria tested, including the slow growing mycobacterium, Mycobacterium bovis BCG. The sensitivity of C2FDG in intact, viable BCG was similar to that of o-nitrophenyl-β-d-galactopyranoside in cell lysates when used to measure lacZ reporter gene activity. C2FDG was approximately 70-fold more sensitive than green fluorescent protein (GFP) in BCG when assayed in a fluorescence plate reader, and comparable to GFP when measured by flow cytometry. These assays provide an important new alternative for the rapid measurement of reporter gene expression in viable bacteria.Keywords
This publication has 22 references indexed in Scilit:
- Identification of Mycobacterium paratuberculosis gene expression signalsMicrobiology, 1997
- FACS-optimized mutants of the green fluorescent protein (GFP)Gene, 1996
- Activity of mycobacterial promoters during intracellular and extracellular growthMicrobiology, 1995
- Flow cytometric study of differentiating cultures of Bacillus subtilisCytometry, 1995
- New vectors for the in vitro generation of alkaline phosphatase fusions to proteins encoded by G + C-rich dnaGene, 1995
- The application of lucifferase as a reporter of environmental regulation of gene expression in mycobacteriaLetters in Applied Microbiology, 1994
- Expression of Escherichia coli ?-galactosidase in Mycobacterium bovis BCG using an expression system isolated from Mycobacterium paratuberculosis which induced humoral and cellular immune responsesMolecular Microbiology, 1992
- Characterization of the micro‐environment of Salmonella typhimurium–containing vacuoles within MDCK epithelial cellsMolecular Microbiology, 1992
- Identification of expression signals of the mycobacteriophages Bxb1, L1 and TM4 using the Escherichia-Mycobacterium shuttle plasmids pYUB75 and pYUB76 designed to create translational fusions to the lacZ geneJournal of General Microbiology, 1992
- The genetic control and cytoplasmic expression of “Inducibility” in the synthesis of β-galactosidase by E. coliJournal of Molecular Biology, 1959