Table 1.
Strains and plasmids used in this study
| 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::Pspac-MCS-lacI lacImc, Cmr |
[23] |
| 8325-4ΔrecU |
NCTC8325-4 recU mutant lacking initial 165 codons |
This study |
| 8325-4recUspaL |
NCTC8325-4 Δspa::Pspac-recU-lacI |
This study |
| BCBRP001 |
NCTC8325-4 ΔrecU Δspa::Pspac-recU-lacI |
This study |
| 8325-4recUi |
NCTC8325-4 ΔrecU Δspa::Pspac-recU-lacI lacImc, 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 Pspac-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 Pspac-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 |
lacImc – cells expressing multicopies of the lacI gene (encoded by pMGPII).