Table 2 Bacterial strains and plasmids used in this study
From: Efficient whole-cell-catalyzing cellulose saccharification using engineered Clostridium thermocellum
Strains/plasmids | Relevant characteristic | Sources |
|---|---|---|
Strains | ||
E. coli | ||
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. thermocellum | ||
DSM1313 | LQ8, wild type stain | DSMZ |
ΔpyrF | Derived from DSM1313, with deleted pyrF gene | This work |
ΔpyrF::CaBglA | Derived from ΔpyrF, expressing CelS-bearing CaBglA | This work |
ΔpyrF::CaBglAm | Derived from ΔpyrF, expressing CelS-bearing CaBglAm | This work |
∆pyrF::pHK-CtBglA-Doc | Derived from ΔpyrF, containing plasmid pHK-CtBglA-Doc | This work |
∆pyrF::pHK-CaBglA-Doc | Derived from ΔpyrF, containing plasmid pHK-CaBglA-Doc | This work |
Plasmids | ||
pHK | pNW33N derivative, E. coli–C. thermocellum shuttle vector, CmR/TmR | [51] |
pHK-∆pyrF | pHK derivative, containing upstream and downstream regions of pyrF for pyrF deletion | This work |
pHK-HR | pHK derivative, containing pyrF cassette, tdk cassette, three regions of homology, and the target DNA sequence to knock in | This work |
pHK-HR-CaBglA | pHK-HR derivative for markerless knock-in of caBglA in the chromosome of DSM 1313 | This work |
pHK-HR-CtBglA | pHK-HR derivative for markerless knock-in of ctBglA in the chromosome of DSM 1313 | This work |
pHK-HR-CaBglAm | pHK-HR derivative for markerless knock-in of caBglAm in the chromosome of DSM 1313 | This work |
pHK-CtBglA-Doc | pHK-HR-CtBglA derivative for expression of CtBglA-Doc in DSM 1313, with the promoter and signal peptide region of CelS | This work |
pHK-CaBglA-Doc | pHK-HR-CaBglA derivative for expression of CaBglA-Doc in DSM 1313, with the promoter and signal peptide region of CelS | This work |
pET28aNS | Expression vector with N-terminal hexahistidine affinity tag, with modified multiple cloning sites | [49] |
pET28aNS-CaBglA | pET28aNS derivative for expression of CaBglA | This work |
pET28aNS-CtBglA | pET28aNS derivative for expression of CtBglA | This work |
pET28aNS-CglT | pET28aNS derivative for expression of CglT | This work |
pET28aNS-Td2f2 | pET28aNS derivative for expression of Td2f2 | This work |