Table 4 Hosts strains, plasmids and primers
Name | Description, relevant genotype or primer sequence (5′ → 3′) | Reference |
|---|---|---|
E. coli host strains | ||
HST08 | F-, endA1, supE44, thi-1, recA1, relA1, gyrA96, phoA, Φ80d lacZ ΔM15, Δ(lacZYA – argF) U169, Δ(mrr – hsdRMS – mcrBC), ΔmcrA, λ– | Takara Bio, Shiga, Japan |
MC4100 | F-, [araD139]B/r, Del(argF-lac)169, λ–-, e14-, flhD5301, Δ(fruK-yeiR)725(fruA25), relA1, rpsL150(strR), rbsR22, Del(fimB-fimE)632(::IS1), deoC1 | [32] |
BW25141 | F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), Δ(phoB-phoR)580, λ-, galU95, ΔuidA3::pir+, recA1, endA9(del-ins)::FRT, rph-1, Δ(rhaD-rhaB)568, hsdR514 | [33] |
BW25113 | F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaD-rhaB)568, hsdR514 | [33] |
WT-∆adhE | adhE null mutant of BW25113 | This study |
WT-∆ldhA | ldhA null mutant of BW25113 | This study |
WT-∆pta | pta null mutant of BW25113 | This study |
Plasmids | ||
pCP20 | FLP+, λ cI857+, λ pR Rep(pSC101 ori)ts, ApR, CmR | [34] |
pKD46 | RepA101ts, ApR, araC-ParaB::gam-bet-exo | [33] |
pK184 | p15A ori, KmR, P lac ::lacZ’ | [35] |
pBBR1MCS-3 | broad host range ori, TcR, P lac ::lacZ’ | [36] |
pUC19 | ColE1 ori, ApR, P lac ::lacZ’ | Invitrogen, Corp., Carlsbad, CA |
pK-scpAKB | From pK184, P lac :: sbm-ygfD-ygfG | This study |
pK-scpAB | From pK184, P lac :: sbm-ygfG | This study |
pB-sucCD | From pBBR1MCS-3, araC-P araB ::sucCD | This study |
pU-adhE(EC) | From pUC19, P lac ::adhE(EC) | This study |
pU-adhE2(CA) | From pUC19, P lac ::adhE2(CA) | This study |
pU-adhE1(CA) | From pUC19, P lac ::adhE1(CA) | This study |
pU-adhEMUT(EC) | From pUC19, P lac ::adhE Glu568Lys(EC) | This study |
pU-adhP(EC) | From pUC19, P lac ::adhP(EC) | This study |
pU-yqhD(EC) | From pUC19, P lac ::yqhD(EC) | This study |
Primers | Â | Â |
v-adhE | AATCTTGCTTACGCCACCTGGAAGTG; CGAACGGTCGCATGAGCAGAAAGCG | This study |
v-pta | GGCATGAGCGTTGACGCAATCAACA; GATCCTGAGGTTAATCCTTCAAACG | This study |
v-ldhA | TCATCAGCAGCGTCAACGGC; ATCGCTGGTCACGGGCTTACCGTT | This study |
m-adhE | CATCCGGAAACTCACTTCGAAAAGCTGGCGCTG; CAGCGCCAGCTTTTCGAAGTGAGTTTCCGGA | This study |
c-scpAB | CCATGATTACGAATTC GCAACAGCTTGCCAACAAGGA; TACCGAGCTCGAATTC TTAATGACCAACGAAATTAGGTTTA | This study |
c-argK | GCTCTAGA ATGTCTTATCAGTATGTTAAGG; GCTCTAGA TTAATCATGATGCTGGC | This study |
c-paraB | CCGCTCTAGA TATTTAGAAAAATAAACAAATAGGGGTTCC; TTGTTTTGCCTGATATTCATGTAAGTTCATTTTTTATAACCTCCTTAGAGCTCGAATTCC | This study |
c-sucCD | ATGAACTTACATGAATATCAGGCAAAACAA; CCCCCCTCGAG TTATTTCAGAACAGTTTTCAGTGCTTCACC | This study |
c-adhE(EC) | CGACTCTAGAGGATCC CATGGCTGTTACTAATGTCGCTGAAC; CTCGGTACCCGGGGATCGATCGGTCAACTAATCCTTAACTGATCG | This study |
c-adhE2(CA) | CGACTCTAGAGGATCC CATGAAAGTTACAAATCAAAAAGAACTAAAACAAAAGC; CTCGGTACCCGGGGATCATAGTCTATGTGCTTCATGAAGCTAATATAATGAAGCAAA | This study |
c-adhE1(CA) | CGACTCTAGAGGATCC CATGAAAGTCACAACAGTAAAGGAATTAGATGAAAA; CTCGGTACCCGGGGATCTTAAGGTTGTTTTTTAAAACAATTTATATACATTTCTTTTATC | This study |
c-adhP(EC) | CGACTCTAGAGGATCC CATGAAGGCTGCAGTTGTTACGAAGG; CTCGGTACCCGGGGATCTTAGTGACGGAAATCAATCACCATGC | This study |
c-yqhD(CA) | CGACTCTAGAGGATCC CATGAACAACTTTAATCTGCACACCC; CTCGGTACCCGGGGATCTTAGCGGGCGGCTTCGTATATACGG | This study |
c-bdhB(CA) | CGACTCTAGAGGATCC CGTGGTTGATTTCGAATATTCAATACCAACTAGAAT; CTCGGTACCCGGGGATCTTACACAGATTTTTTGAATATTTGTAGGACTTCGGA | This study |