Table 5 Bacterial strains, plasmids and primers used in this study
From: Metabolic engineering of Escherichia coli for production of (2S,3S)-butane-2,3-diol from glucose
Name | Characteristic | References |
|---|---|---|
Strains | ||
E. coli DH5α | F− φ80 lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rk−, mk+) supE44λ-thi-1 | Novagen |
E. coli BL21(DE3) | F− ompT gal dcm lon hsdSB(r −B m −B ) λ(DE3) | Novagen |
E. cloacae subsp. dissolvens SDM | Wild type | [5] |
E. coli BL21 (pET28a–lysR–Pabc–budB–budC) | E. coli BL21(DE3) harboring pET28a–lysR–Pabc–budB–budC | This study |
E. coli BL21 (pETDuet–PT7–budB–PT7–budC) | E. coli BL21(DE3) harboring pETDuet–PT7–budB–PT7–budC | This study |
Plasmids | ||
pET28a | Expression vector, Kanr | Novagen |
pETDute-1 | Expression vector, Ampr | Novagen |
pEasy-Blunt | Kanr Ampr oripUC | Transgen |
pET28a–lysR–Pabc–budB–budC | lysR, Pabc, budB and budC of E. cloacae subsp. dissolvens SDM were ligated and cloned into the multiple clone site of pET28a | This study |
pETDuet–PT7–budB–PT7–budC | budB and budC from Enterobacter cloacae subsp. dissolvens SDM were cloned into the two multiple clone sites of pETDuet-1 | This study |
Primers | ||
budB-F (BglII) | 5′-AGATCTAGTGAACAGTGATAAACAG-3′ | This study |
budB-R (XhoI) | 5′-CTCGAGTCACAAAATCTGGCTGAGA-3′ | This study |
budC-F (EcoRI) | 5′-GAATTCAATGCAAAAAGTTGCTCTCG-3′ | This study |
budC-R (HindIII) | 5′-AAGCTTTTAATTGAATACCATCCCACCGT-3′ | This study |
lysR–P abc -F (BglII) | 5′-CGGTAGATCTCTACTCCTCGCTTATCATCG-3′ | This study |
lysR–P abc -R | 5′-CTCACTGTTCATGCTCGTCCTCTTC-3′ | This study |
budB–budC-F | 5′-GAAGAGGACGAGCATGAACAGTGAG-3′ | This study |
budB–budC-R (HindIII) | 5′-GCCTAAGCTTTTAGTTGAACACCATCCCA-3′ | This study |