Table 1 Bacterial strains and plasmids used in this study

Strains

DH5α

f80dlacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(r _k −, m _k +), phoA, supE44, l−, thi-1, gyrA96, relA1

Transgen

BL21(DE3)

ompT gal dcm lon hsdS _B (r _B − m _B −) l (DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5])

Transgen

C. cellulolyticum

H10

ATCC35319, wild type stain

ATCC

H10ΔpyrF

Derived from H10, Ccel_0614 disrupted via homologous recombination

[11]

H10::MspI297s

Derived from H10, mspI ⁻, Ccel_2866:: MspI297s

[14]

H10ΔpyrF::CipC117a

Derived from H10ΔpyrF, Ccel_0728::CipC117a

[11]

H10ΔpyrFΔmspI

Derived from H10ΔpyrF, Ccel_2866::MspI297s

This work

H10ΔpyrFΔcipC

Derived from H10ΔpyrF, Ccel_0728 ::CipC117a

This work

Plasmids

pMTC6

Mls^R, Amp^R, E. coli-C. cellulolyticum shuttle vector, containing PpFbFPm, thl promoter, thl terminator

[14]

pPTK-PpFbFPm

Derived from pMTC6, containing ptk promoter instead of thl promoter

pARA-PpFbFPm

Derived from pMTC6, containing ARAi-PpFbFPm cassette

This work

pGusA2-2tetO1

pIMP1 containing Pcm-2tetO1 promoter and gusA gene

[20]

pARA-GusA

Derived from pARA-PpFbFPm, containing ARAi-gusA cassette

This work

pARA-MazE

Derived from pARA-PpFbFPm, containing T7-mazE cassette

This work

pARA-MazE/F

Derived from pARA-MazE, containing ARAi-mazF cassette

This work

pSY6-mspI

Mls^R, Amp^R, E. coli-C. cellulolyticum shuttle vector, ptb promoter, containing mspI targetron

[14]

pARA-mspI

Derived from pSY6-mspI, containing ARAi system instead of ptb promoter

This work

pARA-PyrF-mspI

Derived from pARA-mspI, containing P _fd -PyrF expression cassette

This work

pGZ-cipC

Mls^R, Amp^R, E. coli-C. cellulolyticum shuttle vector, ptb promoter, containing cipC targetron

[11]

pARA-cipC

Derived from pGZ-cipC, containing ARAi system instead of ptb promoter

This work

pARA-PyrF-cipC

Derived from pARA-cipC, containing P _fd -PyrF expression cassette

This work