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Table 2 Strains, plasmids, transformants, and oligonucleotide primers used in this study

From: Hyper secretion of Thermobifida fusca β-glucosidase via a Tat-dependent signal peptide using Streptomyces lividans

Strain, plasmid, primer, or transformant

Relevant features

Source or reference

Strain

  

Escherichia coli strains

  

Nova blue

endA1 hsdR17(r K12 -m K12 +) supE44 thi-I gyrA96 relA1 lac

Novagene

 

recA1/F’[proAB + lacIq ZΔM15::Tn10(Tetr)]

 

Streptomyces lividans 1326

WT strain

NBRC

Plasmids

  

pUC702-pro-sig-term

Versatile vector for protein expression; thiostrepton resistance marker

[15]

pUC702-plds-bgl

Vector for secreting β-glucosidase(Tfu0937) using plds; thiostrepton resistance marker

[15]

pUC702-xcs-bgl

Vector for secreting β-glucosidase(Tfu0937) using xcs; thiostrepton resistance marker

This study

pCU702-tfu0620s-bgl

Vector for secreting β-glucosidase(Tfu0937) using tfu0620s; thiostrepton resistance marker

This study

pUC702-tfu2486s-bgl

Vector for secreting β-glucosidase(Tfu0937) using tfu2486s; thiostrepton resistance marker

This study

pUC702-tfu2176s-bgl

Vector for secreting β-glucosidase(Tfu0937) using tfu2716s; thiostrepton resistance marker

This study

pUC702-tfu1612s-bgl

Vector for secreting β-glucosidase(Tfu0937) using tfu1612s; thiostrepton resistance marker

This study

pUC702-tfu2788s-bgl

Vector for secreting β-glucosidase(Tfu0937) using tfu2788s; thiostrepton resistance marker

This study

pTYM18

Intergeneric conjugation vector; kanamycin resistance marker

[23]

pTYM18-xcs-bgl

Vector for secreting β-glucosidase(Tfu0937) using xcs; kanamycin resistance marker

This study

pTYM18-plds-bgl

Vector for secreting β-glucosidase(Tfu0937) using xcs; kanamycin resistance marker

This study

Transformants

  

S. lividans/pU-plds-bgl

Transformant harboring pUC702-plds-β-glucosidase (Tfu0937)

[15]

S. lividans/pU-xcs-bgl

Transformant harboring pUC702-xcs-β-glucosidase (Tfu0937)

This study

S. lividans/pU-0620 s-bgl

Transformant harboring pUC702-tfu0620s-β-glucosidase (Tfu0937)

This study

S. lividans/pU-2486 s-bgl

Transformant harboring pUC702-tfu2486s-β-glucosidase (Tfu0937)

This study

S. lividans/pU-2176 s-bgl

Transformant harboring pUC702-tfu2176s-β-glucosidase (Tfu0937)

This study

S. lividans/pU-1612 s-bgl

Transformant harboring pUC702-tfu1612s-β-glucosidase (Tfu0937)

This study

S. lividans/pU-2788 s-bgl

Transformant harboring pUC702-tfu2788s-β-glucosidase (Tfu0937)

This study

S. lividans/pT-xcs-bgl

Transformant integrated pTYM18-xcs-β-glucosidase (Tfu0937)

This study

S. lividans/pTxcs-pUxcs

Transformant integrated pTYM18-xcs-β-glucosidase (Tfu0937) and harboring pUC702-xcs-β-glucosidase (Tfu0937)

This study

S. lividans/pTxcs-pUplds

Transformant integrated pTYM18-xcs-β-glucosidase (Tfu0937) and harboring pUC702-plds-β-glucosidase (Tfu0937)

This study

Oligonucleotide primers

  

xcs_Fw

aGCATGCatgcagcaggacggcacacagcagg

 

xcs_Rv

tttGCTAGCggcgtgggctgtgccgggcagcagc

 

tfu0620s_Fw

TCGTTTAAGGATGCAatgagtaaagttcgtgccacgaaca

 

tfu0620s_Rv

CGATTGCGAGGTCACggcggcgttggccggagcagcgaac

 

tfu2486s_Fw

TCGTTTAAGGATGCAatgtttcgacgtctgcctgtgctgg

 

tfu2486s_Rv

CGATTGCGAGGTCACtgccgcctgagcgtccacgtcagcg

 

tfu2176s_Fw

TCGTTTAAGGATGCAatgtccgtcactgaacctcctcccc

 

tfu2176s_Rv

CGATTGCGAGGTCACttcggcgtgggcggttcccgtggcc

 

tfu1612s_Fw

TCGTTTAAGGATGCAatgacagcaacagcacagcgaacac

 

tfu1612s_Rv

CGATTGCGAGGTCACggcggcagcggagtggacgaggccg

 

Tfu2788s_Fw

TCGTTTAAGGATGCAatgtccgtcactgaacctcctcccc

 

Tfu2788s_Rv

CGATTGCGAGGTCACttcggcgtgggcggttcccgtggcc