Table 3 Bacterial strains and plasmids used in this study

C. salexigens strains

   DSM 3043^T

Wild type

[19]

   CHR61

Spontaneous Rf^r mutant of C. salexigens DSM 3043

[21]

   CHR95

CHR61 ΔeupRmntR::Tn1732; Rf^r Km^r

This study

   CHR161

CHR61 mntR::Ω; Rf^r Sm^r Spcr

This study

   CHR183

CHR61 eupR::Ω; Rf^r Gn^r

This study

E. coli strain

   DH5α

supE44 Δ(lac)U169 ϕ80dlacZ ΔM15 hsdR17 recA1 endA1 gyrA96 thi-1 relA1; host for DNA manipulations

[65]

Plasmids

   pKS(-)

Cloning vector; Ap^r

Stratagene

   pHP45Ω

pBR322 derivative carrying the Ω cassette; Ap^r Smr Sp^r

[50]

   pHP45Ωaac

pBR322 derivative carrying the Ωaac cassette; Ap^r Gm^r Gn^r

[51]

   pRK600

Helper plasmid; Cm^r tra

[66]

   pJQ200-SK

Suicide vector; Gm^r mob sac

[52]

   pSUP102-Gm::Tn1732

Mutagenesis plasmid carrying Tn1732; Cm^r Km^r Gmr

[40, 49]

   pRR1

pKS derivative carrying a 20.8-kb sac I fragment from CHR95 including Tn1732; Ap^r Km^r

This study

   pMntREupR

3-kb Xba I-Apa I fragment from C. salexigens genome (containing orf1, eupR, mntR, orf4) cloned into pKS; Ap^r

This study

   pHpaIMntr

pMntREupR derivative containing a Hpa I recognition site within mntR; Ap^r

This study

   pHindIIIEupR

pMntREupR derivative containing a Hin dIII recognition site within eupR; Ap^r

This study

   PΩMntR

pHpaIMntr derivative with Ω cassette within mntR; Ap^r Sm^r Sp^r

This study

   pΩEupR

pHindIIIEupR derivative with Ωaac cassette within eupR; Ap^r Gn^r Gm^r

This study

   pJQMntR

5-kb Xba I-Apa I fragment from pΩMntR (containing orf1, eupR, mntR::Ω, orf4) cloned into pJQ200-SK; Gm^r Sm^r Sp^r

This study

   pJQEupR

5-kb Xba I-Apa I fragment from pΩEupR (containing orf1, eupR::Ωaac, mntR, orf4) cloned into pJQ200-SK; Gm^r Gn^r

This study