Assessment of GFP fluorescence in cells of Streptococcus gordonii under conditions of low pH and low oxygen concentration

Microbiology
M C HansenS Molin

Abstract

Use of green fluorescent protein (GFP) as a molecular reporter is restricted by several environmental factors, such as its requirement for oxygen in the development of the fluorophore, and its poor fluorescence at low pH. There are conflicting data on these limitations, however, and systematic studies to assess the importance of these factors for growing bacterial cultures are lacking. In the present study, homogeneous expression of the gfpmut3* gene directed by a synthetic constitutive lactococcal promoter was demonstrated in batch cultures and in biofilms of Streptococcus gordonii DL1. A lower limit of oxygen concentration for maturation of the GFP fluorophore was determined: fluorescence was emitted at 0.1 p.p.m. dissolved oxygen (in conventionally prepared anaerobic media lacking reducing agents), whereas no fluorescence was detected in the presence of 0.025 p.p.m. dissolved oxygen (obtained by addition of L-cysteine as reducing agent). When an anaerobically grown (non-fluorescent) >50 microm thick biofilm was shifted to aerobic conditions, fluorescence could be detected within 4 min, reaching a maximum over the next 16 min. It was not possible to detect any fluorescence gradients (lateral or vertical) within the >50 microm...Continue Reading

References

Sep 1, 1991·Research in Microbiology·S D EhrlichA Gruss
Aug 1, 1983·Infection and Immunity·D PerryH K Kuramitsu
Dec 20, 1994·Proceedings of the National Academy of Sciences of the United States of America·R HeimR Y Tsien
Feb 23, 1995·Nature·M P Kilgard, M M Merzenich
Feb 23, 1995·Nature·R HeimR Y Tsien
Sep 5, 1994·FEBS Letters·S Inouye, F I Tsuji
Jun 1, 1993·Journal of Bacteriology·P E Kolenbrander, J London
Nov 1, 1995·Trends in Biochemical Sciences·A B CubittR Y Tsien
Dec 1, 1996·Current Biology : CB·K R SiemeringJ Haseloff
Mar 6, 1997·Biochemical and Biophysical Research Communications·Y KimataK Kohno
Jun 3, 1997·Biochemistry·B G Reid, G C Flynn
Jan 22, 1998·Applied and Environmental Microbiology·P R Jensen, K Hammer
Feb 3, 1998·Biochemical and Biophysical Research Communications·C ZhangP Nagley
Jun 3, 1998·Applied and Environmental Microbiology·J B AndersenS Molin
Mar 1, 1996·Nature Biotechnology·A CrameriW P Stemmer
Feb 1, 1995·Bio/technology·S DelagraveD C Youvan
Apr 2, 1999·Molecular & General Genetics : MGG·A BubertW Goebel
Jan 5, 2000·Applied and Environmental Microbiology·M C GeoffroyA Mercenier
Feb 17, 2000·FEMS Microbiology Letters·P Fernández de PalenciaP López
May 10, 2000·Journal of Microbiological Methods·M C HansenD C White

❮ Previous
Next ❯

Citations

Apr 1, 2008·Expert Opinion on Medical Diagnostics·Alyona Sukhanova, Igor Nabiev
Dec 24, 2015·Frontiers in Microbiology·Anouchka FievetCorinne Aubert
Dec 15, 2015·Parasitology·Victoria Morin-Adeline, Jan Šlapeta
Aug 19, 2007·Journal of Microbiological Methods·Robert J Palmer
Aug 4, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Janne LehtinenEsa-Matti Lilius
Dec 1, 2005·Biotechnology and Bioengineering·Andrés F González BarriosThomas K Wood
Jan 18, 2008·European Journal of Immunology·Ronald BackerJoke M M den Haan
Nov 10, 2010·Environmental Microbiology·Qun MaThomas K Wood
Jun 12, 2013·Environmental Microbiology·Boo Shan TsengMatthew R Parsek
May 12, 2012·ChemSusChem·Ashley E FranksDerek R Lovley
Jan 25, 2011·Microbial Biotechnology·Seok Hoon HongThomas K Wood
Mar 19, 2014·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Yasuo YoshidaFuminobu Yoshimura
Apr 21, 2006·Journal of Microbiological Methods·Chi-Ming Wu, Tung-Ching Chung
Apr 10, 2015·Molecular Microbiology·Nicholas S JakubovicsAlexander H Rickard
Jul 2, 2015·Applied Microbiology and Biotechnology·José M LandeteJuan L Arqués
Oct 20, 2010·FEMS Microbiology Reviews·Nuria AndreuSiouxsie Wiles
May 21, 2010·FEMS Immunology and Medical Microbiology·Ophélie NicolleMartine Bonnaure-Mallet
May 12, 2010·FEMS Microbiology Letters·Michela LizierFranco Lucchini
Jul 7, 2011·PloS One·Simone AymannsBarbara Spellerberg
Jun 6, 2016·World Journal of Microbiology & Biotechnology·José M LandeteJuan L Arqués
Jan 16, 2013·Journal of the American Chemical Society·Chang Kyu ByunShuichi Takayama

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biofilm & Infectious Disease

Biofilm formation is a key virulence factor for a wide range of microorganisms that cause chronic infections.Here is the latest research on biofilm and infectious diseases.