Table 1 Strains and plasmids used in this study
From: Biocatalytic production of the antibiotic aurachin D in Escherichia coli
Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
E. coli strains | ||
DH5α | F– φ80lacZΔ M15 Δ (lacZYA-argF) U169 recA1 endA1 hsdR17 (rK– mK+) phoA supE44 λ- thi–1 gyrA96 relA1 | Invitrogen |
TOP10 | F-mcrA Δ( mrr-hsdRMS-mcrBC) Φ80LacZΔM15 Δ LacX74 recA1 araD139 Δ( araleu) 7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen |
BL21(DE3) | F − ompT hsdSB(rB− mB−) gal dcm (DE3) | Merck KGaA |
S. erecta strains | ||
PD e32 | natural aurachin producer | DSMZ |
Plasmids | ||
pJet1.2/blunt | ampR, E. coli cloning vector (pMB1 ori, eco47IR, PlacUV5, PT7) | Thermo Fisher |
pET-28a(+) | kanR, T7-based E. coli expression vector (pBR322 ori, f1 ori, lacI, rob, PT7, lac operator, His-Tag, T7 terminator) | Novagen |
pSK39 | pET-28a(+) derivative for auaA expression | This study |
pFAB3913 | kanR, E. coli BCD-based expression vector (p15A ori, Ptrc*BCD9, mRFP1) | Mutalik et al. 2013 |
pWaldo-GFPe-SH1446 | kanR, pWaldo-GFPe derivative encoding the POT family transporter SH1446 with a C-terminal TEV-GFP-His-Tag (pBR322 ori, f1 ori, lacI, rob, PT7, lac operator, SH1446, T7 terminator) | Minhas et al. 2018 |
pSK40 | pFAB3913 derivative containing the TEV-folding reporter gfp-His-Tag instead of mRFP1 | This study |
pSK41 | pET-28a(+) derivative harboring the expression cassette of pSK41 instead of the T7 expression cassette (Ptrc* BCD9, TEV-folding reporter gfp-His-Tag) | This study |
pSK42 | pSK41 derivative without lacI | This study |
pSK43 | pSK44 derivative without BCD9 element for monocistronic expression of auaA | This study |
pSK44 | pSK42 derivative for expression of auaA fused to TEV-folding reporter gfp-His-Tag | This study |
pSK45 | pSK42 derivative for expression of auaA_Se fused to TEV-folding reporter gfp-His-Tag | This study |
pSK46 | pSK42 derivative for expression of auaA_Sa fused to TEV-folding reporter gfp-His-Tag | This study |
pJBEI-2997 | cmR, E. coli plasmid harboring the mevalonate pathway (p15A ori, lacI, PlacUV5, atoB, HMGS, tHMGR, MK, PMK, PMD, idi, ispA) | Peralta-Yahya et al. 2011 |