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Table 1 Bacterial strains and plasmids used in this study

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

Strain or plasmid Description Reference or source
Strains
 E. coli
  DH5α Host strain used for general subcloning [61]
  MC1061 Host strain used for subcloning with Lc. lactis derived pNZ8048 vector; recA positive strain Mobitec, Göttingen, Germany
 Lactobacillus plantarum
  423 Probiotic with multiple adhesion genes and a plantaricin bacteriocin producer; forms part of the entiro™ probiotic; originally isolated from sorghum beer Cipla Medpro (Pty.) Ltd.
  423 pNZmazFnisRK Contains the pNZmazFnisRK plasmid; CmR This study
 Enterococcus mundtii
  ST4SA Probiotic with multiple adhesion genes and a mundticin bacteriocin producer; forms part of the entiro™ probiotic; originally isolated from soybeans Cipla Medpro (Pty.) Ltd.
  ST4SA pNZmazFnisRK Contains the pNZmazFnisRK plasmid; CmR This study
 Lactococcus lactis
  pNZ9000 Standard host strain for nisin regulated gene expression; harbors the nisR and nisK nisin regulatory genes integrated into the pepN gene locus Mobitec, Göttingen, Germany
  pNZ9000 pNZmazF Contains the pNZmazF plasmid; CmR This study
Plasmids
 pNZ8048 Broad-host range vector; E. coli shuttle vector; LAB expression vector containing nisin A inducible-promoter (PnisA); CmR Mobitec, Göttingen, Germany; [32]
 pGKV223D E. coli/LAB shuttle vector; LAB expression vector with L23 promoter; EmR University of Gröningen, The Netherlands
 pNZnisRK pNZ8048 vector carrying the nisR and nisK regulatory genes for cloning in LAB strains that do not have the regulatory genes integrated onto the chromosome; CmR This study
 pNZmazF pNZ8048 vector carrying the E. coli mazF toxin gene under the control of the PnisA promoter; CmR This study
 pNZmazFnisRK pNZnisRK vector carrying the E. coli mazF toxin gene under the control of the PnisA promoter; CmR This study
 pNZmazFnisRKerm pNZ8048nisRK vector carrying the E. coli mazF toxin gene under the control of the PnisA promoter and the erm gene; CmR, EmR This study
 pBluescriptKS PCR cloning vector; AmpR Stratagene, California, USA.
 pKSFRT pBluescriptKS plasmid carrying two flippase (FLP) recombination target sequences (FRT) in a direct repeat orientation; AmpR This study
 pKSFRTErm pBluesriptKS plasmid carrying the erm gene flanked by two FRT sequences in a direct repeat orientation; EmR, AmpR This study
 pGKVPldhFLP pGKV223D vector carrying the flippase (FLP) gene downstream of the constitutive L. plantarum 423 lactate dehydrogenase gene Pldh promoter; EmR This study
 pNZasRNA_repAnisRK pNZmazFnisRK vector carrying a 350 bp asRNA_repA transcript downstream of the nisin-inducible PnisA promoter; CmR This study
 pNZasFLPasRNA_repA pNZasRNA_repAnisRK vector carrying the flippase (FLP) gene downstream of the constitutive L. plantarum 423 lactate dehydrogenase gene Pldh promoter; CmR This study
  1. CmR: chloramphenicol resistance; EmR: erythromycin resistance; AmpR: ampicillin resistance