Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages

ABSTRACT Staphylococcus aureus is a prominent global nosocomial and community-acquired bacterial pathogen. A strong restriction barrier presents a major hurdle for the introduction of recombinant DNA into clinical isolates of S. aureus. Here, we describe the construction and characterization of the...

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Autores principales:	Ian R. Monk, Jai J. Tree, Benjamin P. Howden, Timothy P. Stinear, Timothy J. Foster
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2015
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/3901f1788e2a49aab82d50460d7f3a65
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spelling	oai:doaj.org-article:3901f1788e2a49aab82d50460d7f3a652021-11-15T15:49:02ZComplete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages10.1128/mBio.00308-152150-7511https://doaj.org/article/3901f1788e2a49aab82d50460d7f3a652015-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00308-15https://doaj.org/toc/2150-7511ABSTRACT Staphylococcus aureus is a prominent global nosocomial and community-acquired bacterial pathogen. A strong restriction barrier presents a major hurdle for the introduction of recombinant DNA into clinical isolates of S. aureus. Here, we describe the construction and characterization of the IMXXB series of Escherichia coli strains that mimic the type I adenine methylation profiles of S. aureus clonal complexes 1, 8, 30, and ST93. The IMXXB strains enable direct, high-efficiency transformation and streamlined genetic manipulation of major S. aureus lineages. IMPORTANCE The genetic manipulation of clinical S. aureus isolates has been hampered due to the presence of restriction modification barriers that detect and subsequently degrade inappropriately methylated DNA. Current methods allow the introduction of plasmid DNA into a limited subset of S. aureus strains at high efficiency after passage of plasmid DNA through the restriction-negative, modification-proficient strain RN4220. Here, we have constructed and validated a suite of E. coli strains that mimic the adenine methylation profiles of different clonal complexes and show high-efficiency plasmid DNA transfer. The ability to bypass RN4220 will reduce the cost and time involved for plasmid transfer into S. aureus. The IMXXB series of E. coli strains should expedite the process of mutant construction in diverse genetic backgrounds and allow the application of new techniques to the genetic manipulation of S. aureus.Ian R. MonkJai J. TreeBenjamin P. HowdenTimothy P. StinearTimothy J. FosterAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 6, Iss 3 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Ian R. Monk Jai J. Tree Benjamin P. Howden Timothy P. Stinear Timothy J. Foster Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages
description	ABSTRACT Staphylococcus aureus is a prominent global nosocomial and community-acquired bacterial pathogen. A strong restriction barrier presents a major hurdle for the introduction of recombinant DNA into clinical isolates of S. aureus. Here, we describe the construction and characterization of the IMXXB series of Escherichia coli strains that mimic the type I adenine methylation profiles of S. aureus clonal complexes 1, 8, 30, and ST93. The IMXXB strains enable direct, high-efficiency transformation and streamlined genetic manipulation of major S. aureus lineages. IMPORTANCE The genetic manipulation of clinical S. aureus isolates has been hampered due to the presence of restriction modification barriers that detect and subsequently degrade inappropriately methylated DNA. Current methods allow the introduction of plasmid DNA into a limited subset of S. aureus strains at high efficiency after passage of plasmid DNA through the restriction-negative, modification-proficient strain RN4220. Here, we have constructed and validated a suite of E. coli strains that mimic the adenine methylation profiles of different clonal complexes and show high-efficiency plasmid DNA transfer. The ability to bypass RN4220 will reduce the cost and time involved for plasmid transfer into S. aureus. The IMXXB series of E. coli strains should expedite the process of mutant construction in diverse genetic backgrounds and allow the application of new techniques to the genetic manipulation of S. aureus.
format	article
author	Ian R. Monk Jai J. Tree Benjamin P. Howden Timothy P. Stinear Timothy J. Foster
author_facet	Ian R. Monk Jai J. Tree Benjamin P. Howden Timothy P. Stinear Timothy J. Foster
author_sort	Ian R. Monk
title	Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages
title_short	Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages
title_full	Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages
title_fullStr	Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages
title_full_unstemmed	Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages
title_sort	complete bypass of restriction systems for major <named-content content-type="genus-species">staphylococcus aureus</named-content> lineages
publisher	American Society for Microbiology
publishDate	2015
url	https://doaj.org/article/3901f1788e2a49aab82d50460d7f3a65
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Complete Bypass of Restriction Systems for Major <named-content content-type="genus-species">Staphylococcus aureus</named-content> Lineages

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