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Table 1 Strains, plasmids, and primers used in this study

From: Functional heterologous expression of an engineered full length CipA from Clostridium thermocellum in Thermoanaerobacterium saccharolyticum

Strain

Description and characteristics

Reference

T. sacch. M1442

High titer ethanol producing strain

Lee et al., 2011 [47]

T. sacch. DHC6

T. sacch ΔxynA::Clo1313_2747

This study

T. sacch. DHC15

T. sacch ΔxynA::cipA* (wild type sec tag)

This study

T. sacch. DHC16

T. sacch ΔxynA::cipA* (with a deletion in the sec tag)

This study

C. therm. DSM 1313

C. therm wild type obtained from DSMZ culture collection

DSMZ

C. therm. DS11

C. therm 1313 ΔcipA

Olson et al. 2013 [48]

E. coli TOP 10

Chemically competent

Invitrogen

S. cerevisiae FY2

Uracil Auxotroph used for homologous recombination

Winston et. al, 1995 [49]

Plasmid

Description and characteristics

Reference

pMC200

Deletes xynA and replaces it with a kanamycin resistance gene and a removable marker

This study

pMC212

Replaces xynA with Clo1313_2747, a kanamycin resistance gene, and a removable marker

This study

pMC213

Replaces xynA with cipA*, a kanamycin resistance gene, and a removable marker

This study

pMC223

Replaces xynA with Δsec tag cipA*, a kanamycin resistance gene, and a removable marker

This study

Primer

Sequence

Reference

xynA_RT_F

TACTTCAGGATGGGTTGGAACAGG

This study

xynA_RT_R

TCCAATTAGCTGTTCTCCCTGTCG

This study

gamA_RT_F

ATATTCACCAGCAACGCTGGCTTC

This study

gamA_RT_R

AATAAGCCTTTGCCAGTTGTCCGC

This study

Clo1313_2747_RT_F

AGTGGCGTTATACAAACGCTCCTG

This study

Clo1313_2747_RT_R

ATACAACGGAAGCAACGGCAGAAC

This study

cipA*_RT_F

ACGACTATCTTTGCCGCTATGATCCC

This study

cipA*_RT_R

ACTTTAGATCCTACGGCAGCAGTGAC

This study

pMC200_up_F

gtctttcgactgagcctttcgttttatttgatgcctggatcttttctggcctttaatggcg

This study

pMC200_up_R

cagctgaagcttcccggggatcctctagagaattcgagctctcttacttcctccctcagtaaatttaatttattg

This study

pMC200_down_F

ctagataggggtcccgagcgcctacgaggaatttgtatcgaaaaaacaaataatctttaagtaaaaaggcagagagg

This study

pMC200_down_R

ccgtcagtagctgaacaggagggacagctgatagaagtcaaatgcgacaaaaaaacgcc

This study