Stable Expression of Modified Green Fluorescent Protein in Group B Streptococci To Enable Visualization in Experimental Systems

Applied and Environmental Microbiology
Matthew J Sullivan, Glen C Ulett

Abstract

Group B streptococcus (GBS) is a Gram-positive bacterium associated with various diseases in humans and animals. Many studies have examined GBS physiology, virulence, and microbe-host interactions using diverse imaging approaches, including fluorescence microscopy. Strategies to label and visualize GBS using fluorescence biomarkers have been limited to antibody-based methods or nonspecific stains that bind DNA or protein; an effective plasmid-based system to label GBS with a fluorescence biomarker would represent a useful visualization tool. In this study, we developed and validated a green fluorescent protein (GFP)-variant-expressing plasmid, pGU2664, which can be applied as a marker to visualize GBS in experimental studies. The synthetic constitutively active CP25 promoter drives strong and stable expression of the GFPmut3 biomarker in GBS strains carrying pGU2664. GBS maintains GFPmut3 activity at different phases of growth. The application of fluorescence polarization enables easy discrimination of GBS GFPmut3 activity from the autofluorescence of culture media commonly used to grow GBS. Differential interference contrast microscopy, in combination with epifluorescence microscopy to detect GFPmut3 in GBS, enabled visualizat...Continue Reading

References

Dec 1, 1995·Infection and Immunity·S TakahashiY Okuwaki
Nov 15, 1997·Applied and Environmental Microbiology·G V BloembergR Kolter
Dec 11, 1997·Infection and Immunity·V NizetC E Rubens
Jan 22, 1998·Applied and Environmental Microbiology·P R Jensen, K Hammer
May 20, 1999·Journal of Microbiological Methods·D ErrampalliJ T Trevors
Oct 6, 1999·Current Opinion in Microbiology·I MoszerA Danchin
Mar 11, 2000·European Journal of Biochemistry·O ScholzM Niederweis
Aug 23, 2001·Infectious Diseases in Obstetrics and Gynecology·J M SmithS H Jackman
Sep 5, 2002·Molecular Microbiology·Carolyn M Southward, Michael G Surette
Dec 14, 2002·Microbiology·Gregory J TyrrellRichard K Sherburne
Dec 6, 2003·Genome Research·Gila Lithwick, Hanah Margalit
Dec 9, 2003·Applied and Environmental Microbiology·Karyn L NewmanSteven E Lindow
Jul 13, 2004·Trends in Biotechnology·Claes GustafssonJeremy Minshull
Apr 7, 2005·Applied and Environmental Microbiology·Gerald D BaldridgeUlrike G Munderloh
Jul 20, 2005·European Journal of Obstetrics, Gynecology, and Reproductive Biology·Arijaan W Valkenburg-van den BergP Joep Dörr
Aug 19, 2005·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Morven S Edwards, Carol J Baker
Nov 22, 2005·Nature Methods·Nathan C ShanerRoger Y Tsien
Nov 22, 2005·Nature Methods·Jeff W Lichtman, José-Angel Conchello
Nov 22, 2005·Nature Methods·José-Angel Conchello, Jeff W Lichtman
Dec 22, 2005·Nature Biotechnology·Jean-Denis PédelacqGeoffrey S Waldo
Aug 25, 2006·Acta Obstetricia Et Gynecologica Scandinavica·Anouk E MullerP Joep Dörr
Apr 5, 2007·Nature Protocols·Kyoung-Hee Choi, Herbert P Schweizer
Oct 30, 2007·Applied and Environmental Microbiology·Michelle CroninDouwe van Sinderen
Mar 4, 2008·Acta Biomaterialia·James R HullDavid G Castner
Aug 13, 2008·The Journal of Infectious Diseases·Alfredo PezzicoliMarco Soriani
Oct 20, 2010·FEMS Microbiology Reviews·Nuria AndreuSiouxsie Wiles
Jul 7, 2011·PloS One·Simone AymannsBarbara Spellerberg
Oct 19, 2011·Journal of Biomedicine & Biotechnology·Benjamin L DuellGlen C Ulett
Jun 6, 2012·The Veterinary Clinics of North America. Food Animal Practice·Greg Keefe

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.