Table 1 Bacterial strains, plasmids, and oligonucleotides used in this study

E. coli

DH5α

Φ₈₀ dLacZΔM15 recA1 endA1 gyrA96 thi-1 hsdR17(r_k^- m_k⁺) supE44 relA1 deoRΔ(lacZYA-argF)U169

Promega

S. aureus

RN4220

r_k^- m_k⁺; accepts foreign DNA

[20]

RN6390

Prophage-cured wild-type strain

[21]

Newman

Wild-type clinical isolate

[22]

H803

Newman sirA::Km; Km^R

[30]

H1665

Newman Δsfa::Km; Km^R

[9]

H1666

Newman Δsbn::Tet Δsfa::Km; Tet^R Km^R

[9]

H1262

Newman Δhts::Tet; Tet^R

[9]

H1497

Newman sirA::Km Δhts::Tet; Tet^R Km^R

[9]

H2131

Newman sbnA::Tc ΔsfaABCsfaD::Km

This study

H1718

Newman sbnB::Tc ΔsfaABCsfaD::Km

This study

Plasmids

pACYC184

E. coli cloning vector; Cm^R

ATCC

pALC2073

E. coli/S. aureus shuttle vector; Ap^R Cm^R

[26]

pAUL-A

Temperature-sensitive S. aureus suicide vector; Em^R Lc^R

[25]

pDG1514

pMTL23 derivative carrying tetracycline resistance cassette; Ap^R

[24]

pFB5

pALC2073 derivative carrying sbnA; Cm^R

This study

pSED52

pALC2073 derivative carrying sbnB; Cm^R

This study

Oligonucleotides*

Cloning of sbnA into pBC SK+

sbnA5'-SacI

5' TGAGCTCGATTCTGTAGGGCAAACACC 3'

sbnA3'-KpnI

5' TTGGTACCTCTAAGTAACGATCGCCTCG 3'

Amplification/cloning of a tetracycline resistance cassette from pDG1513

Tet5'-NsiI

5' TTGTATATGCATACGGATTTTATGACCGATGA 3'

Tet3'

5' TGTGTGGAATTGTGAGCGGATAAC 3'

Cloning of sbnA into pALC2073

sbnA5'-XhoI

5' TTTCTCGAGATTTTAAATTTGAGGAGGAA 3'

sbnA3'-EcoRI

5' TTTGAATTCCCACATAAACTTGTGAATGATT 3'

Cloning of sbnB into pACYC184

sbnB5'-BamHI

5' TTGGATCCTAGTTTATTCAGATACATGG 3'

sbnB3'-BamHI

5' TTGGATCCTGTCCCAATATTTTGTTGTT 3'

Cloning of sbnB into pALC2073

sbnB5'-EcoRI

5' TTGAATTCTCAAGTGATCCATGTAGATG 3'

sbnB3'-EcoRI

5' TTGAATTCCAATTCCGGCTATATCTTCA 3'