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

S. aureus strain

   RN4220

8325-4 derivative that accept foreign DNA

[55]

   R cls1

RN4220 ΔSA1155; Cm^r

This study

   R cls2

RN4220 ΔSA1891; Tet^r

This study

   R cls1/cls2

RN4220 ΔSA1155/ ΔSA1891; Cm^r Tet^r

This study

   N315

pre-methicillin resistant strain

[56]

   N cls1

N315 ΔSA1155; Cm^r

This study

   N cls2

N315 ΔSA1891; Tet^r

This study

   N cls1/cls2

N315 ΔSA1155/ ΔSA1891; Cm^r Tet^r

This study

   8325-4

NCTC 8325 cured of prophages, rsbU(-)

[57]

   8 cls1

8325-4 ΔSA1155; Cm^r

This study

   8 cls2

8325-4 ΔSA1891; Tet^r

This study

   8 cls1/cls2

8325-4 ΔSA1155/ ΔSA1891; Cm^r Tet^r

This study

   SH1000

8325-4 derivative, rsbU repaired, rsbU(+)

[58]

   S cls1

SH1000 ΔSA1155; Cm^r

This study

   S cls2

SH1000 ΔSA1891; Tet^r

This study

   S cls1/cls2

SH1000 ΔSA1155/ ΔSA1891; Cm^r Tet^r

This study

   MT01

isolate from healthy host, able to generate L-form

This study

   MT01 cls1

MT01 ΔSA1155; Cm^r

This study

   MT01 cls2

MT01 ΔSA1891; Tet^r

This study

   MT01 cls1/cls2

MT01 ΔSA1155/ ΔSA1891; Cm^r Tet^r

This study

plasmids

   pMAD

thermosensitive, pE194^ts-based delivery vector

[53]

   pMADcat1155

pMAD-based SA1155 targeting vector

This study

   pMADtet1891

pMAD-based SA1891 targeting vector

This study

   pHY300PLK

shuttle vector, tet (Tet^R)

Takara, Japan

   pRIT5H

shuttle vector, cm(Cm^R)

[54]

primers

5'-3'

   Tet-F(Sal)

CATATTGTCGAC TAAGTGATGAAATACTG

   Tet-R(EcoRI)

GGAATTC CTGTTATAAAAAAAGGATCAAT

   CAT(Sal)F

CGAGTCGAC GATAAAGTGGGATATTTT

   CAT(Eco)R

CGAATTC CGGGGCAGGTTAGTGACATT

   clsU1p

TGGATCC TGATATTGCTTACATACT

   clsU2p

TGGGTCGAC AAAAAGTACAAATAGC

   clsD1p

CACAGATCT TATGGACTTTAGAAGTT

   clsD2p

TTTAGATCT CAATATCATCCAAATTA

   1891U1

CGGATCC AATAGTCCGACGATAGCT

   1891U2

GAAGTCGAC GTCCTAATAGTAAGTA

   1891D1

TGAATTC ACAAAAGCACGTTATGCT

   1891D2

TGAAGATCT AACATCACAACGGCATA