From: Determination of the native features of the exoglucanase Cel48S from Clostridium thermocellum
Strains/plasmids | Relevant characteristic | Sources |
---|---|---|
Strains | ||
 E. coli | ||
  DH5α | f80dlacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rk − , mk +), phoA, supE44, l − , thi-1, gyrA96, relA1 | Transgen |
  BL21(DE3) | ompT gal dcm lon hsdSB(rB − mB −) l (DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | Transgen |
  BL21(DE3)::pET28aNS-rCel48S_CD | Derived from BL21(DE3), carrying plasmid pET28aNS-rCel48S_CD to express the recombinant Cel48S_CD (rCel48S_CD) | This study |
 C. thermocellum | ||
  ATCC27405 | Wild type stain | ATCC |
  DSM1313 | Wild type stain | DSMZ |
  ΔpyrF | Derived from DSM1313, with deleted pyrF gene | [32] |
  ΔpyrF::Cel48S_CD-His12 | Derived from ΔpyrF, expressing the native Cel48S_CD bearing a His12 tag (ctCel48S_CD -His12) | This study |
Plasmids | ||
 pET28aNS | Expression vector with N-terminal hexahistidine affinity tag, with modified multiple cloning sites | [31] |
 pET28aNS-rCel48S_CD | pET28aNS derivative for expression of Cel48S_CD from C. thermocellum ATCC27405 | This study |
 pHK-HR-CaBglA | Markerless genome editing plasmid for DSM 1313 | [32] |
 pHK-HR-His12TAA | pHK-HR-CaBglA derivative for markerless insertion of His12TAA in the chromosome of DSM 1313 | This study |