Table 2 Strains and plasmids used in this study
Strains and plasmids | Relevant genotype or description | Source or reference |
|---|---|---|
Strains | ||
W3110 | Wild type E. coli | Laboratory stock |
DH5α | Host cells for plasmids amplification | Laboratory stock |
BL21 (DE3) | Host cells for plasmids expression | Laboratory stock |
R1 | W3110 ∆putA | [6] |
R2 | W3110 ∆sucA | This study |
R3 | W3110 ∆aceA | This study |
R4 | W3110 ∆sucA∆aceA | This study |
R5 | W3110 ∆putA∆sucA | This study |
R6 | W3110 ∆putA∆aceA | This study |
R7 | W3110 ∆putA∆sucA∆aceA | [24] |
R8 | W3110 ∆putA∆sucCD∆aceA | This study |
W0 | W3110 harboring p1 | This study |
W1 | R1 harboring p1 | This study |
W2 | R2 harboring p1 | This study |
W3 | R3 harboring p1 | This study |
W4 | R4 harboring p1 | This study |
W5 | R5 harboring p1 | This study |
W6 | R6 harboring p1 | This study |
W7 | R7 harboring p1 | This study |
W8 | R8 harboring p1 | This study |
R8/p2 | R8 harboring p2 | This study |
R8/p3 | R8 harboring p3 | This study |
R8/p4 | R8 harboring p4 | This study |
B0 | BL21 (DE3) harboring p2 | This study |
B1 | BL21 (DE3) harboring p5 | This study |
B2 | BL21 (DE3) harboring p6 | This study |
B3 | BL21 (DE3) harboring p7 | This study |
B4 | BL21 (DE3) harboring p8 | This study |
B5 | BL21 (DE3) harboring p9 | This study |
B6 | BL21(DE3) harboring p10 | This study |
B7 | BL21 (DE3) harboring p11 | This study |
B8 | BL21 (DE3) harboring p12 | This study |
B9 | BL21 (DE3) harboring p13 | This study |
R8/p14 | R8 harboring p14 | This study |
R8/p15 | R8 harboring p15 (H0) | This study |
R8/p16 | R8 harboring p16 | This study |
R8/p17 | R8 harboring p17 | This study |
R8/p18 | R8 harboring p18 | This study |
R8/p19 | R8 harboring p19 | This study |
R8/H1 | R8 harboring H1 | This study |
R8/H2 | R8 harboring H2 | This study |
R8/H3 | R8 harboring H3 | This study |
R8/H4 | R8 harboring H4 | This study |
R8/H5 | R8 harboring H5 | This study |
R8/H6 | R8 harboring H6 | This study |
R8/H7 | R8 harboring H7 | This study |
R8/p20 | R8 harboring p20 | This study |
R8/p21 | R8 harboring p21 | This study |
R8/p22 | R8 harboring p22 | This study |
R8/p23 | R8 harboring p23 | This study |
R8/p24 | R8 harboring p24 | This study |
R8/K0 | R8 harboring K0 | This study |
R8/K1 | R8 harboring K1 | This study |
R8/K2 | R8 harboring K2 | This study |
R8/K3 | R8 harboring K3 | This study |
R8/K4 | R8 harboring K4 | This study |
R8/K5 | R8 harboring K5 | This study |
R8/K6 | R8 harboring K6 | This study |
R8/K7 | R8 harboring K7 | This study |
Plasmids | ||
pKD46 | AmpR plasmid with temperature-sensitive replication and arabinose induction of λ-red recombinase | Laboratory stock |
pCP20 | AmpR and CmR plasmid with temperature-sensitive replication and thermal induction of FLP synthesis | Laboratory stock |
pET24a | High copy number vector, ColE1 ori, T7 lac promoter, KmR | Laboratory stock |
pMAL-C2-X | malE gene encoding MBP fusion protein, tac promoter | Laboratory stock |
pGEX-6P-1 | GST fusion protein, tac promoter | Laboratory stock |
pKK223-3 | AmpR, tac promoter | Laboratory stock |
pACYC-Duet-1 | two multiple cloning sites, T7 lac promoter, P15A origin | Laboratory stock |
pET20b | Bacterial vector for expressing proteins in the periplasm, AmpR, T7 promoter | Laboratory stock |
pET39b | Encodes Dsb tag for export and periplasmic folding of target proteins, KmR, T7 promoter | Laboratory stock |
pET43.1a | High-level expression of peptide sequences fused with the 491 aa Nus•Tag™ protein, AmpR, T7 promoter | Laboratory stock |
pET48b | Encodes Trx tag, KmR, T7 promoter | Laboratory stock |
pMD18T-B74A | pMD18T, p4h-proB74-proA | Laboratory stock |
LGOX | pET28a, lgox | Laboratory stock |
p1 | pET24a, Ptrp::p4h-proB74-proA | This study |
p2 | pET24a, Ptrp::p4h | This study |
p3 | pET24a, Ptrp::p4h-proB74 | This study |
p4 | pET24a, Ptrp::p4h-proB74-proA-proC | This study |
p5 | pACYC-Duet-1, Ptrp::p4h | This study |
p6 | pET24a, p4h | This study |
p7 | pKK223, p4h | This study |
p8 | pGEX-6P-1, p4h | This study |
p9 | pMAL-C2-X, p4h | This study |
p10 | pET20b, p4h | This study |
p11 | pET39b, p4h | This study |
p12 | pET43.1a, p4h | This study |
p13 | pET48b, p4h | This study |
p14 | pMAL-C2-X, Ptac::MBP-p4h-proB74-proA-proC | This study |
p15 (H0) | pET24a, Ptrp::MBP-p4h-proB74-proA-proC | This study |
p16 | pACYC-Duet-1, Ptrp::MBP-p4h | This study |
p17 | pACYC-Duet-1, Ptac::MBP-p4h | This study |
p18 | pACYC-Duet-1, Ptrp::proB74-proA-proC | This study |
p19 | pACYC-Duet-1, Ptac::proB74-proA-proC | This study |
H1 | pET24a, Ptrp::MBP-p4h-Ptrp::proB74-proA-proC | This study |
H2 | pET24a, Ptrp::MBP-p4h-Ptac::proB74-proA-proC | This study |
H3 | pET24a, Ptac::MBP-p4h-proB74-proA-proC | This study |
H4 | pET24a, Ptac::MBP-p4h-Ptrp::proB74-proA-proC | This study |
H5 | pET24a, Ptac::MBP-p4h-T7 t-Ptrp::proB74-proA-proC | This study |
H6 | pET24a, Ptac::MBP-p4h-Ptac::proB74-proA-proC | This study |
H7 | pET24a, Ptac::MBP-p4h-T7 t-Ptac::proB74-proA-proC | This study |
p20 | pACYC-Duet-1, Ptrp:: lgox | This study |
p21 | pACYC-Duet-1, Ptac:: lgox | This study |
p22 | pACYC-Duet-1, Ptrp::icd-gltA-acs | This study |
p23 | pACYC-Duet-1, Ptac::icd-gltA-acs | This study |
K0 | pACYC-Duet-1, Ptrp::lgox-icd-gltA-acs | This study |
K1 | pACYC-Duet-1, Ptrp::lgox-Ptrp::icd-gltA-acs | This study |
K2 | pACYC-Duet-1, Ptrp::lgox-Ptac::icd-gltA-acs | This study |
K3 | pACYC-Duet-1, Ptac::lgox-icd-gltA-acs | This study |
K4 | pACYC-Duet-1, Ptac::lgox-Ptrp::icd-gltA-acs | This study |
K5 | pACYC-Duet-1, Ptac::lgox-T7 t-Ptrp::icd-gltA-acs | This study |
K6 | pACYC-Duet-1, Ptac::lgox-Ptac::icd-gltA-acs | This study |
K7 | pACYC-Duet-1, Ptac::lgox-T7 t-Ptac::icd-gltA-acs | This study |
p24 | pET24a, Ptrp::MBP-p4h-proB74-proA-proC-lgox | This study |