TABLE 1.

Bacterial strains and plasmids

Strain(s) or plasmidRelevant characteristicsReference or source
E. coli DH5arecA1 endA1 hsdR1738
S. pyogenes strains
    HSC5Wild type
    TM-GidA1,TM-GidA2, and TM-GidA3Transposon-generated gidA mutantsaThis study
    HSC5SpcChromosomal insertion of pSPC18 downstream of recF in the HSC5 chromosome without disruption of any gene or operon; previously referred to as “Control”9
    HSC5::pKHC100HSC5 mutant with gidA disruption created with pKHC100bThis study
    HSC5::pKHC101HSC5 mutant created with pKHC101; pSPC18 is inserted between gidA and SPy_2184bThis study
    HSC5::pKHC102HSC5 mutant with SPy_2184 disruption created with pKHC102bThis study
    HSC5::pKHC100::pKHC103Insertion of pKHC103 into HSC5::pKHC100; contains an intact gidA geneThis study
    HSC5::pKHC104HSC5 mutant with mnmE disruption created with pKHC104This study
    HSC5::pKHC105HSC5 mutant with SPy_1173 disruption created with pKHC105This study
    HSC5Spc(pRopB-HA)HSC5Spc transformed with pRopB-HA; produces RopB-HAThis study
    HSC5::pKHC100(pRopB-HA)HSC5::pKHC100 transformed with pRopB-HA; produces RopB-HAThis study
    HSC5::pKHC104(pRopB-HA)HSC5::pKHC104 transformed with pRopB-HA; produces RopB-HAThis study
    HSC5Spc(pSPN-HA)HSC5Spc transformed with pSPN-HA; produces SPN-HAThis study
    HSC5::pKHC100(pSPN-HA)HSC5::pKHC100 transformed with pSPN-HA; produces SPN-HAThis study
    HSC5::pKHC104(pSPN-HA)HSC5::pKHC104 transformed with pSPN-HA; produces SPN-HAThis study
    HSC5Spc(pMF-HA)HSC5spc transformed with pMF-HA; produces MF-HAThis study
    HSC5::pKHC100(pMF-HA)HSC5::pKHC100 transformed with pMF-HA; produces MF-HAThis study
    HSC5::pKHC104(pMF-HA)HSC5::pKHC104 transformed with pMF-HA; produces MF-HAThis study
Plasmids
    pMOD-2<MCS>pUC-based vector containing EZ-Tn5<MCS>Epicenter Inc.
    pMOD-2::Tn5<ΩErm>pUC-based vector containing EZ-Tn5<ΩErm>; derived from pMOD-2<MCS>cThis study
    pSPC18pUC18-based streptococcal integration vector; contains aad9 conferring spectinomycin resistance28
    pCIV2pUC18-based streptococcal integration vector; contains aphA3 conferring kanamycin resistance34
    pSPC18::′recFpSPC18 containing a 0.70-kbp C-terminal segment of recF9
    pKHC100pSPC18 containing a 0.70-kbp internal fragment of gidA generated by PCR with primers GidA-f and GidA-rThis study
    pKHC101pSPC18 containing a 0.62-kbp fragment spanning the gidA-SPy_2184 intergenic region generated by PCR with primers Separate-f and Separate-rThis study
    pKHC102pSPC18 containing a 0.73-kbp internal fragment of SPy_2184 generated by PCR with primers SPy2184-f and SPy2184-rThis study
    pKHC103pCIV2 containing a 2.43-kbp DNA fragment including gidA generated by PCR with primers GidACisComp-f and GidACisComp-rThis study
    pKHC104pSPC18 containing 0.52-kbp internal fragment of mnmE generated by PCR with primers MnmE-f and MnmE-rThis study
    pKHC105pSPC18 containing 0.58-kbp internal fragment of SPy_1173 generated by PCR with primers SPy1173-f and SPy1173-rThis study
    pRopB-HApABG5 derivative containing HA-tagged ropB26
    pSPN-HApABG5 derivative containing HA-tagged spn; same as pJOY717
    pMF-HApABG5 derivative containing HA-tagged mf; same as pJOY217
  • a For detailed information, see Fig. 1.

  • b See Fig. 2.

  • c See Fig. 1.