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Table 1 Bacterial strains and plasmids used in this study

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