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Table 1 Strains and plasmids used in this study

From: The Holliday junction resolvase RecU is required for chromosome segregation and DNA damage repair in Staphylococcus aureus

Strain/Plasmid

Relevant characteristics

Source/ Reference

E. coli

  

DH5α

Cloning strain, recA endA1 gyrA96 thi-1 hsdR17 supE44 relA1 ϕφ80 ΔlacZΔM15

Gibco-BRL

S. aureus

  

NCTC8325-4

MSSA strain

R. Novick

BCBHV008

NCTC8325-4Δspa::P spac -MCS-lacI lacI mc, Cmr

[23]

8325-4ΔrecU

NCTC8325-4 recU mutant lacking initial 165 codons

This study

8325-4recUspaL

NCTC8325-4 Δspa::P spac -recU-lacI

This study

BCBRP001

NCTC8325-4 ΔrecU Δspa::P spac -recU-lacI

This study

8325-4recUi

NCTC8325-4 ΔrecU Δspa::P spac -recU-lacI lacI mc, Cmr

This study

BCBHV017

BCBHV008 strain expressing spoIIIE-yfp from the native chromosomal locus, Cmr

This study

BCBRP002

8325-4recUi mutant strain expressing spoIIIE-yfp, Cmr

This study

Plasmids

  

pMAD

E. coli – S. aureus shuttle vector with the bgaB gene encoding a ß-galactosidase and thermosensitive origin of replication for Gram-positive bacteria, Ampr Eryr

[24]

pBCB13

pMAD derivative with P spac -lacI between up- and downstream regions of the spa gene, Ampr Eryr

[25]

pMGPII

Plasmid encoding lacI gene; Cmr

[26]

pMADrecUKO

pMAD derivative used for deletion of the first 165 codons of recU, Ampr Eryr

This study

pBCB13recUspaL

pBCB13 derivative containing P spac -recU-lacI Ampr Eryr

This study

pMUTINYFPKan

Integrative vector for C-termini YFP fusions; Ampr, Kanr

[27]

pBCBHV007

pMUTINYFPKan containing spoIIIE–yfp Ampr, Kanr

This study

pMADspoIIIEyfp

pMAD derivative containing 3’ end spoIIIE –linker-yfp, Ampr Eryr

This study

pBCBHV008

pMAD derivative containing 3’ end spoIIIE–linker-yfp-3’ 64 bp and downstream region of spoIIIE, Ampr Eryr

This study

  1. lacI mc – cells expressing multicopies of the lacI gene (encoded by pMGPII).