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Table 2 Bacterial strains and plasmids used in this study

From: The toxic guardians — multiple toxin-antitoxin systems provide stability, avoid deletions and maintain virulence genes of Pseudomonas syringae virulence plasmids

Strain/plasmid Main featuresa Source or reference
Escherichia coli  
NEB10β Δ(mrr-hsdRMS-mcrB) deoR recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ rpsL nupG New England Biolabs
S17–1 Strain used to transfer pDR1 plasmid [43]
P. syringae pv. savastanoi  
NCPPB 3335 Pathotype strain, isolated from a diseased olive tree; contains three native plasmids (pPsv48A, pPsv48B and pPsv48C) [78]
Psv48ΔAB NCPPB 3335 derivative, cured of pPsv48A and pPsv48B [18]
UPN25.1 UPN827 derivative containing pPsv48CΔ25, a 5.5 kb spontaneous deletion derivative of pPsv48C that spans the RepJ replicon, with no TA systems. [37]
UPN508 Derivative of NCPPB 3335 containing pPsv48A::Tn5-GDYN1 [18]
UPN827 Derivative of Psv48ΔAB containing plasmid pPsv48C::Tn5-GDYN1 [18]
UPN864 UPN827 derivative, which contains plasmid pPsv48CΔ1 This work
UPN912 Plasmidless derivative of strain NCPPB3335; was obtained by curing pPsv48C::sacB from strain UPN1007 [35]
UPN1007 Derivative of Psv48ΔAB containing plasmid pPsv48C::sacB This work
P. syringae pv. syringae  
B728a Plasmidless bean pathogen CuR, RifR, SmR [79]
Plasmids   
pBlueScript II SK E. coli cloning vector; 2.96 kb, AmpR Stratagene
pBAD24 E. coli expression vector, contains a PBAD promoter inducible with arabinose; 4.5 kb, AmpR [76]
pDR1 Delivery vector for Tn5-GDYN1, based on pSUP2021; KmR, GmR, SmR, SpR; confers sucrose-dependent lethality [43]
pGEM-T Easy E. coli cloning vector; 3 kb, AmpR Promega
pJET1.2 E. coli cloning vector 2.9 kb, AmpR Thermo Fisher Scientific
pK18mobsacB Mobilizable cloning vector, confers sucrose-dependent lethality; KmR, SucS [71]
pKMAG E. coli vector derived from pK184, devoid of the Plac promoter and containing the transcriptional terminator and polylinker from pME6041; 2.6 kb, KmR; accession no. KX714576 [37]
pKMAG-C pKMAG containing the minimal RepA-PFP replicon from pPsv48C, replicates in E. coli and in Pseudomonas; 4.3 kb, KmR; accession no. KX714577 [37]
pME6041 Broad host range cloning vector; 5.6 kb, KmR [80]
pPsv48A Virulence plasmid of NCPPB 3335 (accession n° FR820585); with RepA-PFP replicon; 80.1 kb [18]
pPsv48A::Tn5-GDYN1 Plasmid pPsv48A tagged with Tn5-GDYN1 at position 1469 [18]
pPsv48B Native plasmid of NCPPB 3335 (accession no. FR820586); with RepA-PFP replicon; 45.2 kb [18]
pPsv48C Virulence plasmid of NCPPB 3335 (accession no. FR820587); with RepA-PFP and RepJ replicons; 42.1 kb [18]
pPsv48C::sacB pPsv48C derivative containing the KmR-sacB cassette from pK18mobsacB inserted at position 26,916; KmR, SucS This work
pPsv48C::Tn5-GDYN1 pPsv48C containing Tn5-GDYN1 inserted at position 37,036; KmR, GmR, SucS This work
pPsv48CΔ1 Derives from pPsv48C::Tn5GDYN1 by the spontaneous deletion of 8.3 kb (positions 32,807–41,121 of FR820587) mediated by recombination between two copies of MITEPsy2 This work
pPsv48CΔ25 Spontaneous sucrose-resistant deletion derivative from pPsv48C::sacB, generated by a one-ended transposition of IS801; this plasmid is 5.5 kb long, spanning positions 27,019–32,557 of FR820587 [37]
pRK3A pRK415 derivative containing genes PSPSV_A0043, PSPSV_A0032 and PSPSV_A0020 cloned in tandem in this order, each with their own promoter and under the control of the Plac promoter from the vector; TcR This work
pRK3C pRK415 derivative containing genes PSPSV_C0050, PSPSV_C0008 and PSPSV_C0003 cloned in tandem in this order, each with their own promoter and under the control of the Plac promoter from the vector; TcR This work
pRK415 Broad host range cloning vector; 10.5 kb, TcR [81]
  1. aAbbreviations: Amp ampicillin, Cu copper, Km kanamycin, Gm gentamicin, Rif rifampicin, Sm streptomycin, Sp spectinomycin, Suc sucrose, Tc tetracyclin. Superscripts R and S denote resistance or susceptibility, respectively