Table 1 Bacterial strains and plasmids used in the present study
Strain/plasmidGenotype/phenotypeReference
Escherichia coli
CC118 λpiraraD, Δ(ara, leu), ΔlacZ74, phoA20, galK, thi-1, rspE, rpoB, argE, recA1, λpir[40]
DH5αF, ϕ80ΔdlacZM15, Δ(lacZYAargF)U169, endA1, recA1, hsdR17 (rKmK+), deoR, thi-1, supE44, λ, gyrA96, relA1Gibco/BRL
HH26Marker exchange mobilization strain for conjugal transfer[39]
Pectobacterium atrosepticum
PCF80Δcas::cat derivative of SCRI1043[42]
PCF196Δcas1, markerless derivative of SCRI1043This study
SCRI1043Wild type (WT)[61]
Plasmids
pBAD30Arabinose inducible vector, p15a replicon, ApR[44]
pBluescript II KS+Cloning vector, ColE1 replicon, ApRStratagene
pKNG101Suicide vector, sacBR, mobRK2, R6K ori, SmR[36]
pNJ5000Mobilizing plasmid for marker exchange, TcR[39]
pPF170pTRB30 with N-His6-tagged PatCas1, KmR (aka pJSC1)[42]
pPF571pQE-80LoriT-mCherry-derivative, TcR[27]
pPF574pQE-80LoriT-mCherry-derivative, CRISPR1spacer 1 F priming plasmid, TcR[27]
pPF565Δcas1 knockout intermediate, pBluescript II KS+, ApRThis study
pPF568Δcas1 knockout in pKNG101, SmRThis study
pPF626pTRB30 with N-His6-tagged PatCas1 D269A, KmRThis study
pPF692pBAD30 with N-His6-tagged PatCas1, ApR This study
pPF693pBAD30 with N-His6-tagged PatCas1 D269A, ApRThis study
pTRB30pQE-80L (Qiagen) based expression vector, ApR replaced by KmR, KmR[42]