Construction of improved tools for protein localization studies in Streptococcus pneumoniae.

We have constructed a set of plasmids that allow efficient expression of both N- and C-terminal fusions of proteins of interest to fluorescent proteins mCherry, Citrine, CFP and GFP in the Gram-positive pathogen Streptococcus pneumoniae. In order to improve expression of the fluorescent fusions to l...

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Autores principales:	Mafalda X Henriques, Maria João Catalão, Joana Figueiredo, João Paulo Gomes, Sergio R Filipe
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/67e49e3fb30348019514056328010134
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spelling	oai:doaj.org-article:67e49e3fb303480195140563280101342021-11-18T08:00:32ZConstruction of improved tools for protein localization studies in Streptococcus pneumoniae.1932-620310.1371/journal.pone.0055049https://doaj.org/article/67e49e3fb303480195140563280101342013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23349996/?tool=EBIhttps://doaj.org/toc/1932-6203We have constructed a set of plasmids that allow efficient expression of both N- and C-terminal fusions of proteins of interest to fluorescent proteins mCherry, Citrine, CFP and GFP in the Gram-positive pathogen Streptococcus pneumoniae. In order to improve expression of the fluorescent fusions to levels that allow their detection by fluorescence microscopy, we have introduced a 10 amino acid tag, named i-tag, at the N-terminal end of the fluorescent proteins. This caused increased expression due to improved translation efficiency and did not interfere with the protein localization in pneumococcal bacteria. Localizing fluorescent derivatives of FtsZ, Wzd and Wze in dividing bacteria validated the developed tools. The availability of the new plasmids described in this work should greatly facilitate studies of protein localization in an important clinical pathogen.Mafalda X HenriquesMaria João CatalãoJoana FigueiredoJoão Paulo GomesSergio R FilipePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e55049 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mafalda X Henriques Maria João Catalão Joana Figueiredo João Paulo Gomes Sergio R Filipe Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
description	We have constructed a set of plasmids that allow efficient expression of both N- and C-terminal fusions of proteins of interest to fluorescent proteins mCherry, Citrine, CFP and GFP in the Gram-positive pathogen Streptococcus pneumoniae. In order to improve expression of the fluorescent fusions to levels that allow their detection by fluorescence microscopy, we have introduced a 10 amino acid tag, named i-tag, at the N-terminal end of the fluorescent proteins. This caused increased expression due to improved translation efficiency and did not interfere with the protein localization in pneumococcal bacteria. Localizing fluorescent derivatives of FtsZ, Wzd and Wze in dividing bacteria validated the developed tools. The availability of the new plasmids described in this work should greatly facilitate studies of protein localization in an important clinical pathogen.
format	article
author	Mafalda X Henriques Maria João Catalão Joana Figueiredo João Paulo Gomes Sergio R Filipe
author_facet	Mafalda X Henriques Maria João Catalão Joana Figueiredo João Paulo Gomes Sergio R Filipe
author_sort	Mafalda X Henriques
title	Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
title_short	Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
title_full	Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
title_fullStr	Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
title_full_unstemmed	Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
title_sort	construction of improved tools for protein localization studies in streptococcus pneumoniae.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/67e49e3fb30348019514056328010134
work_keys_str_mv	AT mafaldaxhenriques constructionofimprovedtoolsforproteinlocalizationstudiesinstreptococcuspneumoniae AT mariajoaocatalao constructionofimprovedtoolsforproteinlocalizationstudiesinstreptococcuspneumoniae AT joanafigueiredo constructionofimprovedtoolsforproteinlocalizationstudiesinstreptococcuspneumoniae AT joaopaulogomes constructionofimprovedtoolsforproteinlocalizationstudiesinstreptococcuspneumoniae AT sergiorfilipe constructionofimprovedtoolsforproteinlocalizationstudiesinstreptococcuspneumoniae
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Construction of improved tools for protein localization studies in Streptococcus pneumoniae.

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