TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionSource
B. subtilis
    CU741trpC2 leuC748
    LA213trpC2 leuC7 amyE::alfA-lacZ (Cmr)pOF5LA2 × CU741
    WILA2LA213 derivative with alfA preceded by synthetic, wild-type repeat sequencespWILA2 × CU741
    LMLA2WILA2 derivative carrying an alteration in the first repeatpLMLA2 × CU741
    MDLA13WILA2 derivative carrying alterations in the first and second repeatspMDLA13 × CU741
    RMLA11WILA2 derivative carrying an alteration in the third repeatpRMLA1 × CU741
E. coli
    JM103Δlac-pro thi rpsL supE sbcB hsdR4 F′ [traD36 proAB+ lacIq lacZΔM15]51
S. cerevisiae
    AH109MATa trpl-901 leu2-3,112 ura3-52 his3-200 ga14 Δgal80 ΔLYS2:: GAL1UAS-GAL1TATA-HIS3 GAL2]UAS-GAL2TATA-ADE2 URA3::MEL1UAS-MEL1TATA-lacZClontech
Plasmids
    pDH88Integration vector carrying Pspac promoter16
    pDG148Carries Pspac promoter before muticloning sites42
    pQE8Expression vector for construction of His-tagged proteinsQiagen
    pIS284Delivers promoter-E. coli lacZ fusion to B. subtilis amyE locus and carries EcoRI, BamHI and XbaI sites in that order upstream of the lacZ geneI. Smith
    pBET131Wild-type plasmid (Fig. 1A)46
    pBIF2965pBET131 carrying a deletion from the BamHI site to position 2964This study
    pBIC6561pBIF6561 carrying Pspac promoter upstream of position 2965This study
    pBIF6561pBIF2965 carrying a deletion spanning positions 3033 to 3060This study
    pBIAW2pBET131 carrying synthetic, wild-type DNA between BamHI and SnaBI sitesThis study
    pBIBM6pBET131 carrying a deletion between the BamHI and SnaBI sitesThis study
    pBIBM19pBIAW2 carrying nucleotide alterations in the first repeatThis study
    pBIBM24pBIAW2 carrying nucleotide alterations in the second repeatThis study
    pBIBM34pBIAW2 carrying nucleotide alterations in the third repeatThis study
    pBIBM8pBIAW2 carrying nucleotide alterations in both the first and second repeatsThis study
    pDGEN51pDG148 carrying alfA downstream of Pspac promoterThis study
    pENSA51pDGEN51 carrying a blunt-ended SacI site in alfAThis study
    pDGEX561pDG148 carrying alfA and alfB downstream of Pspac promoterThis study
    pEXSA564pDGEX561 carrying a blunt-ended SacI site in alfAThis study
    pOF5LA2pIS284 carrying the promoter region of alfA before lacZThis study
    pWILA2pORF5LA2 derivative carrying synthetic, wild-type repeat sequencesThis study
    pLMLA2pWILA2 derivative carrying an alteration in the first repeatThis study
    pMDLA13pWILA2 derivative carrying alterations in the first two repeat sequencesThis study
    pRMLA1pWILA2 derivative carrying an alteration in the third repeatThis study
    pQE8OF6pQE8 carrying alfBThis study
    pGBKT7Kmr; cloning vector carrying GAL4 DNA-binding domain (BD)Clontech
    pGADT7Ampr; cloning vector carrying GAL4 activation domain (AD)Clontech
    pGBA3pGBKT7 carying a fusion between alfA and GAL4 DNA BDThis study
    pGAB142pGADT7 carrying a fusion between alfB and GAL4 ADThis study
    pGB64pGBKT7 carrying a fusion between alfB and GAL4 DNA BDThis study
    pGA51pGADT7 carrying a fusion between alfA and GAL4 ADThis study
    pGBKT7-53Carrying a fusion between murine p53 and GAL4 DNA BD in pGBKT7Clontech
    pGADT7-TCarrying a fusion between simian virus 40 (SV40) large T antigen and GAL4 AD in pGADT7Clontech