High-level fluorescence labeling of gram-positive pathogens.

Fluorescence labeling of bacterial pathogens has a broad range of interesting applications including the observation of living bacteria within host cells. We constructed a novel vector based on the E. coli streptococcal shuttle plasmid pAT28 that can propagate in numerous bacterial species from diff...

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Autores principales: Simone Aymanns, Stefanie Mauerer, Ger van Zandbergen, Christiane Wolz, Barbara Spellerberg
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Publicado: Public Library of Science (PLoS) 2011
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Acceso en línea:https://doaj.org/article/1b8fd3324d5e4a43930173a5b2b70265
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spelling oai:doaj.org-article:1b8fd3324d5e4a43930173a5b2b702652021-11-18T06:51:37ZHigh-level fluorescence labeling of gram-positive pathogens.1932-620310.1371/journal.pone.0019822https://doaj.org/article/1b8fd3324d5e4a43930173a5b2b702652011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21731607/?tool=EBIhttps://doaj.org/toc/1932-6203Fluorescence labeling of bacterial pathogens has a broad range of interesting applications including the observation of living bacteria within host cells. We constructed a novel vector based on the E. coli streptococcal shuttle plasmid pAT28 that can propagate in numerous bacterial species from different genera. The plasmid harbors a promoterless copy of the green fluorescent variant gene egfp under the control of the CAMP-factor gene (cfb) promoter of Streptococcus agalactiae and was designated pBSU101. Upon transfer of the plasmid into streptococci, the bacteria show a distinct and easily detectable fluorescence using a standard fluorescence microscope and quantification by FACS-analysis demonstrated values that were 10-50 times increased over the respective controls. To assess the suitability of the construct for high efficiency fluorescence labeling in different gram-positive pathogens, numerous species were transformed. We successfully labeled Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus dysgalactiae subsp. equisimilis, Enterococcus faecalis, Enterococcus faecium, Streptococcus mutans, Streptococcus anginosus and Staphylococcus aureus strains utilizing the EGFP reporter plasmid pBSU101. In all of these species the presence of the cfb promoter construct resulted in high-level EGFP expression that could be further increased by growing the streptococcal and enterococcal cultures under high oxygen conditions through continuous aeration.Simone AymannsStefanie MauererGer van ZandbergenChristiane WolzBarbara SpellerbergPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 6, p e19822 (2011)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Simone Aymanns
Stefanie Mauerer
Ger van Zandbergen
Christiane Wolz
Barbara Spellerberg
High-level fluorescence labeling of gram-positive pathogens.
description Fluorescence labeling of bacterial pathogens has a broad range of interesting applications including the observation of living bacteria within host cells. We constructed a novel vector based on the E. coli streptococcal shuttle plasmid pAT28 that can propagate in numerous bacterial species from different genera. The plasmid harbors a promoterless copy of the green fluorescent variant gene egfp under the control of the CAMP-factor gene (cfb) promoter of Streptococcus agalactiae and was designated pBSU101. Upon transfer of the plasmid into streptococci, the bacteria show a distinct and easily detectable fluorescence using a standard fluorescence microscope and quantification by FACS-analysis demonstrated values that were 10-50 times increased over the respective controls. To assess the suitability of the construct for high efficiency fluorescence labeling in different gram-positive pathogens, numerous species were transformed. We successfully labeled Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus dysgalactiae subsp. equisimilis, Enterococcus faecalis, Enterococcus faecium, Streptococcus mutans, Streptococcus anginosus and Staphylococcus aureus strains utilizing the EGFP reporter plasmid pBSU101. In all of these species the presence of the cfb promoter construct resulted in high-level EGFP expression that could be further increased by growing the streptococcal and enterococcal cultures under high oxygen conditions through continuous aeration.
format article
author Simone Aymanns
Stefanie Mauerer
Ger van Zandbergen
Christiane Wolz
Barbara Spellerberg
author_facet Simone Aymanns
Stefanie Mauerer
Ger van Zandbergen
Christiane Wolz
Barbara Spellerberg
author_sort Simone Aymanns
title High-level fluorescence labeling of gram-positive pathogens.
title_short High-level fluorescence labeling of gram-positive pathogens.
title_full High-level fluorescence labeling of gram-positive pathogens.
title_fullStr High-level fluorescence labeling of gram-positive pathogens.
title_full_unstemmed High-level fluorescence labeling of gram-positive pathogens.
title_sort high-level fluorescence labeling of gram-positive pathogens.
publisher Public Library of Science (PLoS)
publishDate 2011
url https://doaj.org/article/1b8fd3324d5e4a43930173a5b2b70265
work_keys_str_mv AT simoneaymanns highlevelfluorescencelabelingofgrampositivepathogens
AT stefaniemauerer highlevelfluorescencelabelingofgrampositivepathogens
AT gervanzandbergen highlevelfluorescencelabelingofgrampositivepathogens
AT christianewolz highlevelfluorescencelabelingofgrampositivepathogens
AT barbaraspellerberg highlevelfluorescencelabelingofgrampositivepathogens
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