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Table 2 List of integration vectors and recombinant LAB strains

From: Development of a novel selection/counter-selection system for chromosomal gene integrations and deletions in lactic acid bacteria

Plasmid Modified strain Locus Upstream region of homology Downstream region of homology Region deleted on chromosome/plasmid Element(s) integrated onto chromosome/plasmid
pNZKOplaA::ErmFfluc L. plantarum 423 plaA::erm-ffluc plaA 902 bp upstream of plaA 69 bp internal fragment of plaA, 330 bp plaB bacteriocin immunity gene and 214 bp of plaC bacteriocin translocation protein 208 bp, including
102 bp of plaA
3204 bp erm-ffluc gene cassette
pNZKOmunA::CatFfluc E. mundtii ST4SA munA::cat-ffluc munA 911 bp upstream of munA, including 135 bp internal fragment of munA ORF 633 bp internal fragment of munB translocation gene 260 bp, including 18 bp of munA ORF 2471 bp cat-ffluc gene cassette
pNZKIaap::FRTerm L. plantarum 423 aap::frt-erm aap 909 bp internal fragment of aap ORF 929 bp internal fragment of aap ORF None 1458 bp FRT-flanked erm gene
pNZKIaap::FRTerm L. plantarum 423 aap::frt_um   909 bp internal fragment of aap ORF 929 bp internal fragment of aap ORF 1063 bp FRT-flanked erm resistance gene excised 335 bp including a 96 bp FRT-scar
pNZKOsrtA::FRTerm E. mundtii ST4SA srtA::frt-erm srtA 1011 bp upstream of srtA, including 57 bp internal fragment of srtA ORF, 333 bp mazG nucleotide pyrophosphohydrolase gene and 270 bp fragment of mfs transporter ORF 1011 bp downstream of srtA, including 57 bp internal fragment of srtA ORF and 576 bp internal fragment of bglG transcriptional anti-terminator ORF 580 bp internal fragment of the srtA ORF 1458 bp FRT-flanked erm gene
pNZKOsrtC::FRTerm E. mundtii ST4SA srtC::frt-erm srtC 1011 bp upstream of srtC, including 59 bp of srtC ORF and 892 bp of EbpCfm pilus subunit protein gene 376 bp downstream of srtA, including 60 bp of srtC ORF and 225 bp internal fragment of hemolysin III ORF 726 bp internal fragment of the srtC ORF 1458 bp FRT-flanked erm gene
  1. The integration vectors and their relevant characteristics are shown, and their construction is described in detail in the main text (pNZKOmunA::CatFfluc, pNZKIaap::FRTerm and pNZKOsrtA::FRTerm), Additional file 1: Text S1 (pNZKOplaA::ErmFfluc and pNZKOsrtC::FRTerm) and Additional file 1: Figs S2–S6. plaA: plantaricin 423 bacteriocin gene; erm: erythromycin resistance gene; ffluc; Photinus pyralis firefly luciferase gene; munA; mundticin ST bacteriocin gene; cat: chloramphenicol resistance gene; aap; FRT: flippase recombination target; L. plantarum 423 adhesion gene; um: unmarked; srtA: E. mundtii ST4SA sortaseA gene; srtC: E. mundtii ST4SA sortaseC gene