Table 1 Bacteria and plasmids used in the study

Escherichia coli

1830

pro¯ met¯ Kan^r Nm^r, containing transposon Tn5 on the suicidal plasmid pBJ4JI

[44]

DH5α

supE44ΔlacU169(Φ80lacZΔM15) hsdR17recA1 gyrA96thi-1relA1

[39]

BL21(DE3)

hsdS gal(λcIts857 ind1 Sam7 nin5 lac UV5-T7 gene 1)

[45]

Pectobacterium carotovorum subsp. carotovorum

3F-3

Pcc, wild-type

Laboratory stock

F-rif-18

3F3, Rif^r, wild-type

This study

TF1-1

F-rif-18, fliC::Tn5, Rif^r, Kan^r

This study

TF 1-2

F-rif-18, CarocinS2::Tn5, Rif^r, Kan^r

This study

SP33

Pcc, wild-type

Laboratory stock

Plasmid

pMCL210

p15A, Cml^r, Low copy number

[46]

pGEM T-Easy

Amp^r; lacZ cloning vector

Promega

pET32a

Amp^r; expression vector with the N-terminal His-tag

Novagen

pET30b

Kan^r; expression vector with the C-terminal His-tag

Novagen

pMS2KI

5.7-kb BamHI DNA fragment harboring carocin S2 gene from 3F3 genome, cloned into pMCL210

This study

pEN2K*

caroS2K subcloned into pET32a

This study

pES2KI

Derived from pEN2K; deleted series of Tag element in front of expressed caroS2K

This study

pEH2KI*

Derived from pES2KI; adding (His)₆-Tag adjacent to caroS2I

This study

pGS2I

caroS2I and its putative promoter from pMS2KI, subcloned into pGEM T-easy

This study

pECS2I*

caroS2I subcloned into pET30b, but the expressed fusion CaroS2I has no activity

This study

pES2I

Derived form pECS2I, the (His)₆-Tag element was deleted

This study