Table 1.

Bacterial strains and plasmids used

Strain or plasmidRelevant characteristic and/or genotypeaComments (reference or source)
Strains
S. aureus
  RN4220Restriction-deficient mutagenized RN450Shuttle plasmid host (16)
  450MCms Ems GmsMcr (HE) Tcs Bla RN450 transformed with COL mec region DNA (1)
  N315PCms Emr GmsMcr (HE) Tcs Bla IntactmecR1-mecI; clinical isolate from Japan (13)
  67-0Mcr Emr (HE) Bla Clinical isolate from California, 1984 (5)
E. coli
  TB1 recA + lacI q lacZΔM15 Host for pUC vectors (26)
  DH5αF recA1 hsdR17φ80dlacZM15Δ(lacZYA-argF) endA1 deoR gyr A96 thi-1 relA supE44 Recombination-deficient host for pUC derivatives (Gibco)
Plasmids
 pUC19Apr; 2.7 kbGeneral-purpose cloning vector (26)
 pGEX-2TApr; 4.8 kbVector for the expression of GST-protein fusions (Pharmacia)
 pSK950Spr Tcr Emr ts; 10.4 kb E. coli-S. aureus shuttle vector for integration of sequences into the S. aureus lipase gene with φL54a att site. The original pCL84 plasmid was modified by addition of pE194 at the PstI site (17).
 pG0630 or pG0631SprTcr Emr ts; 16.4 kbA 2.4-kb fragment from N315P(pG0630) or 67-0(pG063), containing themecR1-mecI intergenic region with divergentmecR1-mecI and mecA promoters, and the first 50 bases of the mecA coding sequence cloned atEcoRI-BamHI sites of pSK950. A 3.3-kb fragment containing promoterless lacZ was then cloned at theBamHI site to generate a mecA-lacZtranscriptional fusion (this study).
  • a Abbreviations: Ap, ampicillin; Bla, β-lactamase; Cm, chloramphenicol; Em, erythromycin; Gm, gentamicin; HE, heterotypic; Mc, methicillin; MCS, multiple cloning site; Sp, spectinomycin; Tc, tetracycline; ts, temperature sensitive.