Table 1.

Bacterial strains, plasmids, and phage

Strain, plasmid, or phageaRelevant characteristic(s)bReference or source
E. coli
 Strains
  JF626Aps, Se8
  M15(pREP4)KmrQiagen
  DH5Aps, SeGibco BRL
  DH10BrecA1 deoR mcrA mcrBC mrr hsdRMSGibco BRL
  MJB1308Aps, seg, carrying pMJB460This work
  MJB1309Aps,sei, carrying pMJB461This work
 Plasmids
  pGEM-7Zf(+)Apr, containsblaPromega
  pGEM-3Zf(+)AprPromega
  pQE-31, pQE-32AprQiagen
  pMJB38Apr, pBR322 containing a 624-bp fragment ofsea8
  pMJB124Apr, pGEM7-Zf(+) with a 1.8-kbp insert that contains sec14
  pMJB460Apr, pGEM7-Zf(+) with a 2.5-kbpHindIII seg-containing fragment from FRI572 inserted into the HindIII site (Fig. 2A)This work
  pMJB461Apr, pGEM7-Zf(+) with a 2.5-kbpHindIII sei-containing fragment from FRI445 inserted into the HindIII site (Fig. 2B)This work
  pMJB464Apr, pGEM7-Zf(+) digested withSmaI/EcoRI and ligated to a 2.1-kbpseg-containing insert obtained from the digestion of pMJB462 with ClaI (in blunted multiple-cloning site)/EcoRIThis work
  pMJB465Apr, pGEM7-Zf(+) digested with HindIII/SmaI and ligated to a 1.73-kbp HindIII/RsaIsei-containing insert obtained by digesting pMJB461 withHindIII/EcoRI followed by a RsaI partial digest (Fig. 2B)This work
  pMJB474pQE-32 with a 800-bp insert containing seg, encodes HisSEG (Fig.2A)This work
  pMJB475pQE-31 with a 770-bp insert containing sei, encodes HisSEI (Fig. 2B)This work
S. aureus
 Strains
  FRI337Sea+ Sed+M. S. Bergdollc
  FRIS6Sea+Seb+M. S. Bergdoll
  FRI578See+M. S. Bergdoll
  FRI400Sec+M. S. Bergdoll
  FRI569EnterotoxigenicdM. S. Bergdoll
  FRI393
  FRI591
  FRI574
  FRI772
  FRI662
  FRI572
  FRI445
  FRI572ContainssegM. S. Bergdoll
  FRI445ContainsseiM. S. Bergdoll
  RN450NCTC 8325 derivative cured of prophages φ11, φ12, and φ13R. P. Novick (53)
  RN4220Se38
  RN7497Contains pI524, recipient strain for pRN5548-derived plasmidsS. J. Projane (56)
  RN8117Cmr Cdr, contains pRN5548 and pI524S. J. Projan (56)
  ISP2073Spa55
  MJB894Sea+, ISP2073 carrying pMJB193Lab strain
  MJB1315Sei+, Spa, ISP2073 carrying pMJB466This work
  MJB1316Seg+ (inducible), RN7497 carrying pMJB467This work
  MJB1317Seg+, RN7497 carrying pMJB468This work
  MJB1318Sei+, Spa, ISP2073 carrying pMJB469This work
  MJB1320Sei+ (inducible), RN7497 carrying pMJB471This work
  MJB1321Sei+, RN7497 carrying pMJB473This work
 Phage 80αGeneralized transducing phage50
 Plasmids
  pC194CmrR. P. Novick (32)
  pRN5548Cmr, blaZpromoter and ribosome-binding site (no start codon) followed by a multiple-cloning siteS. J. Projan (56)
  pI524Cdr, contains β-lactamase control elements49
  pMJB193AprCmr, contains 2.5-kbp HindIII fragment containing sea7
  pMJB462pMJB460 + pC194 ligated in the SmaI site of pGEM7Zf(+), contains segThis work
  pMJB463Cmr, pMJB462 with a translation termination signal introduced into the seg BsmI site by digesting pMJB462 with BsmI, creating blunt ends, and ligating the plasmid to a SMURFT linker (17-mer; Pharmacia Biotech) that contains ochre translation termination signals in all three reading framesThis work
  pMJB466AprCmr, pMJB465 + pC194 joined at HindIII site, contains a 1.73-kbp sei-containing insert (Fig. 2B)This work
  pMJB467Cmr, pRN5548 (digested with PstI/SmaI) ligated to a 1.9-kbp insert containing seg (obtained by isolating the fragment from an NsiI/EcoRV digest of pMJB460) in which seg is transcribed from the inducible S. aureus β-lactamase promoter (Fig. 2A)This work
  pMJB468Cmr, isogenic to pMJB467 except for a translation termination signal present in seg, pRN5548 with a 1.9-kbp seg-containing insert prepared from pMJB463 as described for pMJB467This work
  pMJB469Cmr, pMJB466 with a translation termination signal introduced intosei at the KpnI site (after the first 66 deduced amino acids of SEI) by performing a partial KpnI digest, creating blunt ends, and religating the plasmidThis work
  pMJB470Cmr, pRN5548 with a 1.73-kbpEcoRI/BamHI sei-containing fragment (same genomic fragment as in pMJB465) obtained from pMJB465This work
  pMJB471Cmr, pRN5548 with a 1.24-kbp fragment containing sei behind the β-lactamase promoter (plasmid created by digesting pMJB470 with XbaI to remove DNA 5′ of sei) (Fig. 2B)This work
  pMJB472Cmr, pMJB465 with a translation termination signal introduced into sei at theKpnI site as described for pMJB469This work
  pMJB473Cmr, isogenic to pMJB471 except for a translation termination signal in the KpnI site ofsei, made by digesting pMJB472 withBamHI/EcoRI and then XbaI, isolating the 1.24-kbp fragment, and ligating it to similarly digested pRN5548This work
  pMJB476Cmr, pMJB461 (sei on 2.5-kbp insert) with pC194 inserted into theSmaI site (Fig. 2B)This work
  • a Address strain requests to Rodney Welch, Department of Medical Microbiology and Immunology, University of Wisconsin—Madison, Madison, WI 53706.

  • b Ap, ampicillin; Cm, chloramphenicol; Cd, cadmium; Km, kanamycin; blaZ, β-lactamase; Spa, protein A nonproducer; Sea+, Seb+, Sed+, Seg+, and Sei+, producers of staphylococcal enterotoxin types A, B, D, G, and I, respectively; Se, non-enterotoxin producer.

  • c University of Wisconsin—Madison, Food Research Institute, Madison, Wis.

  • d Culture supernatants produce an emetic response when administered orally to primates.

  • e Wyeth-Ayerst Research, Pearl River, N.Y.