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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
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
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]
  1. Antibiotic resistance genes are abbreviated in the table as follows: amp, ampicillin; tet, tetracycline; cat, chloramphenicol; spec, spectinomycin; and neo, neomycin. SD, Shine-Dalgarno.