TABLE 1.

Bacterial strains and plasmids used in this work

Strain or plasmidGenotypeDesignationSource or reference
Strains
    TOP10E. coli host strain used for recombinant pPL2 plasmids
    DH5αE. coli host strain for recombinant pKSV7 plasmids
    SM10E. coli host strain for conjugation of pPL2 plasmids
    NF-E1254E. coli BL21 Star (DE3) with pNF-E1251 for PrsA2 protein expression and purificationThis work
    NF-L100L. monocytogenes 10403S parent strain2
    NF-L476L. monocytogenes 10403S actA-gus-plcB59
    NF-L116710403S actA-gus-neo prfA(L140F)L140F45
    DP-L216110403S containing in-frame deletion of hly geneΔhly27
    NF-L1456NF-L476 with group II intron (targetron) insertion into lmo2219 (prsA2) at position 457prsA2::TThis work
    NF-L1475NF-L1456 with integrated pPL2-prsA2.prsA2::T + prsA2This work
    NF-L1587NF-L1456 with prsA2::T replaced with a ΔprsA2::erm mutationThis work
    NF-L1635DP-L2161 (Δhly) with ΔprsA2::ermΔprsA2::erm ΔhlyThis work
    NF-L1637NF-L1167 [prfA(L140F) with ΔprsA2::erm]prfA(L140F) ΔprsA2::ermThis work
    NF-L1659NF-L1637 [prfA(L140F) ΔprsA2::erm] with integrated pPL2-prsA2 (pNF1255)prfA(L140F) ΔprsA2::erm + pPL2-prsA2This work
    NF-L165110403S with ΔprsA2::ermΔprsA2::ermThis work
    NF-L1656NF-L1651 (ΔprsA2::erm) with integrated pPL2-prsA2 (pNF1255)ΔprsA2::erm + pPL2-prsA2This work
    HEL 40210403S with prsA1 in-frame deletionΔprsA1This work
Plasmids
    pNF1256pKSV7 with ΔprsA2 SOE fragment with internal SalI restriction siteThis work
    pNF1264pNF1256 with erm resistance cassette inserted at SalI restriction siteThis work
    pHY304Plasmid pHY304 containing constitutive erm cassette26
    pNF1251pET-TOPO N-His with prsA2 open reading frameThis work
    pNF1255pPL2-prsA2 containing prsA2 with 636 bp upstream of start ATG and 68 bp downstream of stop TAAThis work
    pNF1520pNL9146 with targeted targetron group II intron for gene disruption of prsA2 at nucleotide position 457This work