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Table 2 Plasmids and strains used in this study

From: Single mutation at a highly conserved region of chloramphenicol acetyltransferase enables isobutyl acetate production directly from cellulose by Clostridium thermocellum at elevated temperatures

Name

Descriptions

Source

Plasmids

 pNW33N

BacillusE. coli shuttle vector, CmR, pBC1 ori for Gram-positive strains, pBR322 ori for E. coli, source of CATSa

Bacillus Genetic Stock Center

 pETDuet-1

pBR322 ori, AmpR, lacI, T7lac promoter

Novagen

 pET_CATSa

CATSa wild-type encoding gene between BamHI, SacI site, pETDuet-1 backbone, 6× His-tag at N-terminus

This study

 pET_CATSa F97W

F97W site-directed variant, pET_CATSa backbone

This study

 pHS0024

CATSa wild-type gene under C. thermocellum PgapDH promoter, downstream of Clo1313_2927 for the transcription terminator, tdk operon under cbp promoter substituting with the native cat selection marker, pNW33N plasmid backbone

This study

 pHS0024_F97W

CATSa F97W site-directed mutated from pHS0024

This study

Strains

 E. coli Top10

Host for molecular cloning, mcrA, (mrr-hsdRMS-mcrBC), Phi80lacZ(del)M15, ∆lacX74, deoR, recA1, araD139, (ara-leu)7697, galU, galK, rpsL(SmR), endA1, nupG

Invitrogen

 E. coli BL21 (DE3)

E. coli B dcm, ompT, hsdS(rB-mB-), gal

Invitrogen

 M1354

C. thermocellum DSM1313 ∆hpt

[37]

 HSCT0101

M1354 harboring pHS0024

This study

 HSCT0102

M1354 harboring pHS0024_F97W

This study

  1. The plasmids containing mutagenized genes are presented in Additional file 1: Table S1