TABLE 1.

Plasmids and strains used in this study

Strain or plasmidRelevant characteristic(s)Source or reference
Plasmids
    pJDC9Nonreplicative E. coli-S. pneumoniae shuttle vector8
    pAS1pJDC9 derivative30
    pASPB2pAS1 with B2 promoter, cloned in XhoI/EcoRI sitesThis study
    pDSNdepDS derivative39
    pDSNde-6hispDSNde; creates a six-histidine fusion tag39
    pGK13-KANpJDC9 and pGK13 derivative; S. pneumoniae replicative plasmid with kanamycin resistance26
    pNR2pDSNde containing vicR, cloned in NdeI/BamHIThis study
    pvicKpDSNde containing vicK, cloned in NdeI/BamHIThis study
    pKOHK2pAS1 containing the 5′ end of vicK, cloned in XhoI/EcoRIThis study
    pKO423pAS1 containing the 5′ 1,270 nucleotides of vicK, cloned in EcoRI/BamHIThis study
    pKOOX2pAS1 containing the 5′ end of vicX, cloned in XhoI/EcoRIThis study
    pOBRR2pASPB2 containing the 5′ end of vicR plus a ribosomal binding site, cloned in EcoRI and BamHIThis study
    pOBHK2pOBRR2 containing the 5′ end of vicK, cloned in NdeI and BamHIThis study
    pOBOX2pOBRR2 containing the 5′ end of vicX, cloned in NdeI and BamHIThis study
    pHK2KOPpASPB2 containing the 5′ end of vicK, cloned in EcoRI/BamHIThis study
    pMBP-VanSpMAL-c2 containing VanS41
    pMBP-VanRpMAL-c2 containing VanR41
Strains
    S. pneumoniae
        R6S. pneumoniae laboratory strain7
        KNR. 7/87 14.8.91Serotype 4 strain, sequenced by Human Genome Sciences1
        Sp1Serotype 22 strain, clinical isolateRoche strain collection
        Sp1711Serotype 3 strain, clinical isolateRoche strain collection
        vicKKOKnockout of vicK in R6 with pKOHK2This study
        vicXKOKnockout of vicX in R6 with pKOOX2This study
        SOBvicROverexpression of vic operon, obtained with pOBRR2This study
        SOBvicKOverexpression of vicK and vicX, obtained with pOBHK2This study
        SOBvicXOverexpression of vicX, obtained with pOBOX2This study
        vicKKOPOverexpression of vicX, obtained with pHK2KOPThis study
    E. coli
        SURE strainHost strain for E. coli cloning stepsStratagene
        M15[REP4]Qiagen expression host strain39