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

From: Secretory production of tetrameric native full-length streptavidin with thermostability using Streptomyces lividans as a host

Strain, plasmid, primer, or transformant

Relevant features

Source or reference

Strains

  

Escherichia coli

  

Nova blue

endA1 hsdR17(r K12 -m K12 +) supE44 thi-I gyrA96 relA1 lac recA1/F’[proAB+ lacIq ZΔM15::Tn10(Tetr)]

Novagen

S17-1 λpir

TpR SmR recA, thi, pro, hsdR-M+RP4: 2-Tc:Mu: Km Tn7 λpir

BIOMEDAL

BL21 (DE3) pLysS

F– ompT hsdS(r B – m B – ) gal dcm λ(DE3) pLysS (Camr) (λ(DE3): lacI,lacUV5-T7 gene 1,ind1,sam7,nin5)

TAKARA BIO

Streptomyces lividans

  

Streptomyces lividans 1326

WT strain (NBRC 15675)

NBRC

Plasmids

  

pTONA4

Versatile vector for protein expression in Streptomyces; thiostrepton and kanamycin resistance marker

[24]

pUC702-pro-sig-term

Versatile vector for protein expression; thiostrepton resistance marker

[16]

pUC702-ps-Savcore

Vector for Sav (core) expression; thiostrepton resistance marker

This study

pUC702-ps-Savnat

Vector for Sav (native) expression; thiostrepton resistance marker

This study

pTONA4-Savnat

Vector for Sav (native) expression; thiostrepton and kanamycin resistance marker

This study

pTONA4-ps-Savnat

Vector for Sav (native) expression; thiostrepton and kanamycin resistance marker

This study

pTONA4-Savcore

Vector for Sav (core) expression; thiostrepton and kanamycin resistance marker

This study

pTONA4-SavΔC

Vector for Sav-ΔC expression; thiostrepton and kanamycin resistance marker

This study

pTONA4-SavΔN

Vector for Sav-ΔN expression; thiostrepton and kanamycin resistance marker

This study

pWI3SAFlo318

Vector used as a template for amplifying the synthetic gene of streptavidin

[25]

pColdI

Versatile vector for protein expression in E. coli; ampicillin resistance marker

TAKARA BIO

pColdI-Sav

Vector for Sav expression; ampicillin resistance marker

This study

Transformants

  

S. lividans/Savnat

S. lividans transformant harboring pTONA4-Savnat

This study

S. lividans/Savcore

S. lividans transformant harboring pTONA4- Savcore

This study

S. lividans/psSavnat

S. lividans transformant harboring pTONA4-ps-Savnat

This study

S. lividans/SavΔC

S. lividans transformant harboring pTONA4- SavΔC

This study

S. lividans/SavΔN

S. lividans transformant harboring pTONA4- SavΔN

This study

E. coli/SavEco

E. coli transformant harboring pColdI-Sav

This study

Oligonucleotide primers

  

Sav_F

TCGTTTAAGGATGCAatgcgcaagatcgtcgttgca

 

Sav_R

CGCTCAGTCGTCTCAgtggtggtggtggtggtgctgctgaacggcgtcgagcgggtt

 

SavΔN_F

GCCAGCGCTTCGGCAgaggccggcatcaccggcacctgg

 

SavΔC_R

CGCTCAGTCGTCTCAgtggtggtggtggtggtgggaggcggcggacggcttca

 

Sav-sig_F

TCGTTTAAGGATGCAatgcgcaagatcgtcgttgcagccatcgccgtttccctgaccacggtctcgattacggccagcgcttcggca

 

Sav-sig_R

tgccgaagcgctggccgtaatcgagaccgtggtcagggaaacggcgatggctgcaacgacgatcttgcgcatTGCATCCTTAAACGA

 

SavΔN_to_ps_F

GCGGCTCCGGCCTTCgaggccggcatcaccggcacctgg

 

Sav_os_to_ps_F

GCGGCTCCGGCCTTCatgcgcaagatcgtcgttgca

 

ps_F

TCGTTTAAGGATGCAGCATGCTCCGCCACCGGCTCCGCCG

 

Kpn1_Xho1_Stav_F

GGGGTACCCTCGAGGCCGAGGCCGGCATCACCGGCACCTGG

 

Stav_TAG_BamH1_EcoR1_R

GGAATTCGGATCCCGCTAGGAGGCGGCGGACGGCTTCACCTTGGTGAAGGT

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