Table 5 Bacterial strains and plasmids.

E. coli

5-alpha

cloning strain

New England Biolabs

S. aureus

Newman

ATCC25904, methicillin susceptible

[72]

RN4220

NCTC 8325-4-r, restriction negative strain

[73]

COL

methicillin resistant (MRSA)

[60]

RH7657

Newman ΔvraSR::ery

this study

RH7661

Newman ΔvraDE::ery

this study

RH7788

RN4220 ΔvraDE::ery

this study

RH7790

RN4220 ΔvraSR::ery

this study

Plasmids

pGEM-3zf(+)

cloning vector, amp^r (E. coli)

Promega

pKOR1

gene replacement vector, amp^r (E. coli), chl^r (S. aureus)

[69]

pKTH3762

pGEM-3zf(+) containing the vraSR deletion cassette, amp^r (E. coli)

this study

pKTH3763

pKOR1 containing the vraSR deletion cassette, amp^r (E. coli), chl^r ery^r (S. aureus)

this study

pKTH3764

pGEM-3zf(+) containing the vraDE deletion cassette, amp^r (E. coli)

this study

pKTH3765

pKOR1 containing the vraDE deletion cassette, amp^r (E. coli), chl^r ery^r (S. aureus)

this study