Table 3 Bacterial strains and plasmids used or constructed in this study
From: High external pH enables more efficient secretion of alkaline α-amylase AmyK38 by Bacillus subtilis
Strain or plasmid | Relevant properties† | Source or reference |
|---|---|---|
Strain | ||
Bacillus subtilis | ||
168 | trpC2 | [5] |
MGB625 | MGB604 Δ(yeeK-yesX) | [8] |
MGB723 | MGB625 Δ(pdp-rocR) | [8] |
MGB874 | MGB860 Δ(yncM-yndN) | [8] |
MGB625Δr1 | MGB625 Δ(pdp-yxeD)::cat | [11] |
MGB625Δr2 | MGB625 Δ(yxeC-yxnA)::cat | [11] |
MGB625Δr3 | MGB625 Δ(yxaD-fbp)::cat | [11] |
MGB625Δr4 | MGB625 Δ(yydD-rocR)::cat | [11] |
MGB625ΔyydD-yycN | MGB625 Δ(yydD-yycN)::cat | [11] |
MGB625ΔrapG-phrG | MGB625 Δ(rapG-phrG)::cat | [11] |
MGB625ΔrocF-rocR | MGB625 Δ(rocF-rocR)::cat | [11] |
MGB625Δpdp-rocD | MGB625 Δ(pdp-rocD)::cat | This study |
874DEFR | MGB874 cat-rocDEF-rocR was inserted at the Δ(pdp-rocR) locus | [11] |
874RocR | MGB874 cat -rocR was inserted at the Δ(pdp-rocR) locus | This study |
874DEFRΔrocG | 874DEFR ΔrocG::spec | [11] |
874DEFRΔrocABC | 874Δ DEFRΔrocABC::spec | This study |
R-726 | MGB874 ΔaprE::Pspac-rocG spec (pAPNC213-rocG) | This study |
R-1645 | 168 ΔamyE::PhtrB-lacZ cat (pDN110) | This study |
R-1646 | MGB874 ΔamyE::PhtrB-lacZ cat (pDN110) | This study |
R-1926 | MGB874 ΔaprE::Pspac-rocG spec ΔamyE::PhtrB-lacZ cat (pDN110) | This study |
168ΔcssRS | 168 ΔcssRS::spec | This study |
874ΔcssRS | MGB874 ΔcssRS::spec | This study |
168ΔdltB | trpC2 ΔdltB::neo | This study |
874ΔdltB | MGB874 ΔdltB::neo | This study |
Escherichia coli | ||
HB101 | supE44 Δ(mcrC-mrr) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 leuB6 thi-1 | Takara Bio |
Plasmid | ||
pHY300PLK | Shuttle vector for E. coli and B. subtilis | Takara Bio |
pHYK38 | pHY300PLK carrying the gene for AmyK38 from Bacillus sp. strain KSM-K38 fused to the promoter, SD, and leader peptide sequences of egl-237, containing amp and tet | This study |
pAPNC213 | The integration vector containing Pspac promoter into the aprE locus by double crossover using spec | [35] |
pAPNC213-rocG | pAPNC213 containing the SD sequence and coding region of rocG (−25 to 1275, 1325 bp) | This study |
pDL2 | Integration vector for introduction of single-copy transcriptional fusions to lacZ by double crossover at the amyE locus (cat) | [36] |