TABLE 1

Strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s)aReference or source
Strains
    E. coli TOP10Cloning hostThermo Fisher
    E. coli DH5αCloning hostLaboratory stock
    L. lactis Il1403Cloning host60
    L. plantarum WCFS1Host strain12
    L. plantarum NZ3557WCFS1, Δacm2::cat34
    L. plantarum IM167WCFS1, pEV, pSgRNA-notargetThis study
    L. plantarum IM132WCFS1, pSIP-SH-dCas9This study
    L. plantarum IM133WCFS1, pSIP-SH-dCas9, pSgRNA-notargetThis study
    L. plantarum IM134WCFS1, pSIP-SH-dCas9, pSgRNA-acm2This study
    L. plantarum IM137WCFS1, pSIP-SH-dCas9, pSgRNA-dnaAThis study
    L. plantarum IM188WCFS1, pSIP-SH-dCas9, pSgRNA-ezrAThis study
    L. plantarum IM141WCFS1, pSIP-SH-dCas9, pSgRNA-lp_1247 (cozE homolog)This study
    L. plantarum IM142WCFS1, pSIP-SH-dCas9, pSgRNA-lp_2217 (cozE homolog)This study
    L. plantarum IM130WCFS1, pSIP-SH-dCas9, pSgRNA-lp_1247–lp_2217 (cozE homologs)This study
    L. plantarum IM138WCFS1, pSIP-SH-dCas9, pSgRNA-lp_3683 (eloR homolog)This study
    L. plantarum IM139WCFS1, pSIP-SH-dCas9, pSgRNA-lp_1637 (khpA homolog)This study
Plasmids
    pSIP403spp-based expression vector; Emr; 256rep, pUCori; PsppA::gusA15
    pSIP411spp-based expression vector; Emr; SH71rep; PsppQ::gusA16
    pSIPdCas9Emr; 256rep, pUCori; PsppA::dCas9This study
    pSIP-SH-dCas9Emr; SH71rep; PsppA::dCas9This study
    pPEPX-P3-sgRNAluc22
    pValacTemplate for chloramphenicol resistance gene (Cmr)55
    pEVEmr; control plasmid, empty vector61
    pSgRNA-notargetCmr, 256rep, pUCori P3::sgRNA-notargetThis study
    pSgRNA-acm2Cmr 256rep, pUCori P3::sgRNA-lp_2645This study
    pSgRNA-dnaACmr 256rep, pUCori P3::sgRNA-lp_0001This study
    pSgRNA-ezrACmr 256rep, pUCori P3::sgRNA-lp_2328This study
    pSgRNA-lp_1247Cmr 256rep, pUCori P3::sgRNA-lp_1247This study
    pSgRNA-lp_2217Cmr 256rep, pUCori P3::sgRNA-lp_2217This study
    pSgRNA-lp_1247- lp_2217Cmr 256rep, pUCori P3::sgRNA-lp_1247-lp_2217This study
    pSgRNA-lp_3683/eloRCmr 256rep, pUCori P3::sgRNA-lp_3683This study
    pSgRNA-lp_1637/khpACmr 256rep, pUCori P3::sgRNA-lp_1637This study
  • a Em, erythromycin; Cm, chloramphenicol.