Green fluorescent protein as a new expression marker in mycobacteria

Molecular Microbiology
L KremerC Locht

Abstract

This study describes the use and the advantages of the green fluorescent protein (GFP) as a reporter molecule for mycobacteria. The gfp gene from Aequorea victoria was placed under the control of the hsp60 promoter in the shuttle vector pGFM-11. The gfp expression in the recombinant Mycobacterium smegmatis and BCG was readily detected on agar plates by the development of an intense green fluorescence upon irradiation with long-wave u.v. light. In mycobacteria containing a pGFM-11 derivative that lacks the hsp60 promoter, no fluorescence was observed. However, this plasmid was successfully used as a promoter-probe vector to identify BCG promoters. The fluorescence emission of GFP in mycobacteria harbouring pGFM-11 and grown in liquid media could be quantified by spectrofluorimetry. This allowed for easy assessment of drug susceptibility. As GFP does not require the addition of substrates or co-factors, the green fluorescent bacilli could be directly observed within infected macrophages using fluorescence and laser confocal microscopy, or in tissue sections of infected mice. Finally, infected cells or free-living recombinant mycobacteria could also be analysed by flow cytometry. The GFP thus appears to be a convenient reporter fo...Continue Reading

Citations

May 20, 1999·Journal of Microbiological Methods·D ErrampalliJ T Trevors
Sep 18, 2002·Research in Microbiology·James A TriccasWarwick J Britton
Mar 6, 1999·Current Opinion in Microbiology·R H Valdivia, S Falkow
Aug 18, 2000·Immunology and Cell Biology·J A Triccas, B Gicquel
Aug 21, 2002·The Journal of Experimental Medicine·F GeissmannJ Davoust
Jun 30, 2000·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·I Hautefort, J C Hinton
Aug 3, 2002·Microbiology and Immunology·Takashi FunatsuHideo Namiki
Jul 24, 2012·Applied and Environmental Microbiology·Jennifer L EitsonNicolai S C van Oers
Jan 5, 2000·Applied and Environmental Microbiology·M C GeoffroyA Mercenier
Nov 28, 2001·Applied and Environmental Microbiology·J S WebbG D Robson
Nov 5, 2005·Applied and Environmental Microbiology·Salme TimmuskE Gerhart H Wagner
Dec 2, 2005·Journal of Molecular Microbiology and Biotechnology·Agné KulytéLiam Good
Jul 25, 2009·Medicinal Research Reviews·Arvind S NegiSuman P S Khanuja
Mar 22, 2005·Microbes and Infection·Rajinder P S PartiRanjana Srivastava
Dec 22, 2005·International Journal of Antimicrobial Agents·Saiko ObataTeruo Kirikae
Oct 3, 2014·BioMed Research International·Verónica Ambriz-AviñaMario Pedraza-Reyes
Sep 5, 2015·Journal of Immunology Research·Piotr SzpakowskiMagdalena Kowalewicz-Kulbat
Feb 6, 2007·International Immunopharmacology·Salvatore Pasquale PreteOrnella Franzese
Mar 31, 2010·BMC Microbiology·Frédéric-Antoine DauchyJeanne Maugein
Oct 7, 2003·Annual Review of Microbiology·Josephine E Clark-Curtiss, Shelley E Haydel

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antifungals

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Antitubercular Agents (ASM)

Antitubercular agents are pharmacologic agents for treatment of tuberculosis. Discover the latest research on antitubercular agents here.

Addiction

This feed focuses mechanisms underlying addiction and addictive behaviour including heroin and opium dependence, alcohol intoxication, gambling, and tobacco addiction.

Bacterial Respiration

This feed focuses on cellular respiration in bacteria, known as bacterial respiration. Discover the latest research here.

Antifungals (ASM)

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Antitubercular Agents

Antitubercular agents are pharmacologic agents for treatment of tuberculosis. Discover the latest research on antitubercular agents here.