TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or descriptiona | Source or referenceb |
|---|---|---|
| Strains | ||
| B. subtilis | ||
| B15029 | Wild type (Ent+) | DSM15029 |
| B6633 | Wild type (Sub+) | ATCC 6633 |
| B6633.MB1 | ΔspaS amyE::PspaS-lacZ (Specr Cmr Sub−) | 26 |
| B15029.TSp01 | ΔetnS (Specr Ent−) | This work |
| B15029.TSp03 | ΔetnS amyE::PspaS-spaS (Specr Neor) | This work |
| B15029.TSp05 | ΔetnS amyE::PspaS-spaS(W1I) (Specr Neor) | This work |
| B15029.TSp10 | ΔetnS amyE::PspaS-spaS(W1I/K2T) (Specr Neor) | This work |
| B15029.TSp11 | ΔetnS amyE::PspaS-spaS(W1I/K2T/E4I) (Specr Neor) | This work |
| B15029.TSp24 | ΔetnS amyE::PspaS-spaS(W1I/K2T/E4I/V12K) (Specr Neor) | This work |
| B15029.TSp32 | ΔetnS amyE::PspaS-spaS(W1I/K2T/E4I/V12K/Q17 M) (Specr Neor) | This work |
| B15029.TSp57 | ΔetnS amyE::PspaS-spaS(W1E) (Specr Neor) | This work |
| B15029.TSp58 | ΔetnS amyE::PspaS-spaS(W1Y) (Specr Neor) | This work |
| B15029.TSp59 | ΔetnS amyE::PspaS-spaS(W1F) (Specr Neor) | This work |
| B15029.TSp60 | ΔetnS amyE::PspaS-spaS(W1G) (Specr Neor) | This work |
| B15029.TSp66 | ΔetnS amyE::PspaS-spaS(W1A) (Specr Neor) | This work |
| B15029.TSp67 | ΔetnS amyE::PspaS-spaS(W1D) (Specr Neor) | This work |
| B15029.TSp68 | ΔetnS amyE::PspaS-spaS(K2T/E4I/V12K/Q17M) (Specr Neor) | This work |
| B15029.TSp69 | ΔetnS amyE::PspaS-spaS(W1H) (Specr Neor) | This work |
| B15029.TSp71 | ΔetnS amyE::PspaS-spaS(W1K) (Specr Neor) | This work |
| B15029.TSp72 | ΔetnS amyE::PspaS-spaS(W1R) (Specr Neor) | This work |
| B15029.TSp81 | ΔetnS amyE::PspaS-spaS(W1L) (Specr Neor) | This work |
| B15029.TSp82 | ΔetnS amyE::PspaS-spaS(W1V) (Specr Neor) | This work |
| B15029.TSp89 | amyE::PspaS-spaS(W1E) (Neor) | This work |
| B15029.TSp90 | amyE::PspaS-spaS(W1D) (Neor) | This work |
| B15029.TSp91 | amyE::PspaS-spaS(W1A) (Neor) | This work |
| B15029.TSp98 | amyE::PspaS-spaS(W1K) (Neor) | This work |
| L. lactis | ||
| NZ9800 | NZ9700 derivative ΔnisA | 39 |
| NZ9800.AUT1 | NZ9800 containing pNZ8148 PnisA-lacZ (Cmr) | This work |
| E. coli DH5α | recA1 endA1 gyrA96 thi hsdR17(rK− mK+) relA1 supE44 ϕ80ΔlacZΔM15 Δ(lacZYA-argF)U169 | Laboratory stock |
| Kocuria rhizophila K9341 | Test strain for MIC determination | ATCC 9341 |
| S. cerevisiae CEN.PK2 | MATa/α ura3-52/ura3-52 trp1-289/trp1-289 leu2-3,112/leu2-3,112 his3Δ1/his3Δ1 MAL2-8c/MAL2-8c SUC2/SUC2 | 34 |
| Plasmids | ||
| pRS416 | bla URA3 shuttle vector | 33 |
| ECE74 | pCm::Spc | 35 |
| pMLK83 | amyE integrative plasmid (BGSC accession no. ECE103) | 36 |
| pSD27 | pMLK83 removal of gusA by EcoRI | This work |
| pRS426 | bla URA3 shuttle vector | 37 |
| pTSp10 | bla etnS′ Specr ′etnS | This work |
| pTSp16 | bla amyE′ PspaS-spaS Neor ′amyE | This work |
| pTSp25 | bla amyE′ PspaS-spaS(W1I) Neor ′amyE | This work |
| pTSp33 | bla amyE′ PspaS-spaS(W1I/K2T) Neor ′amyE | This work |
| pTSp34 | bla amyE′ PspaS-spaS(W1I/K2T/E4I) Neor ′amyE | This work |
| pTSp45 | bla amyE′ PspaS-spaS(W1I/K2T/E4I/V12K) Neor ′amyE | This work |
| pTSp49 | bla amyE′ PspaS-spaS(W1I/K2T/E4I/V12K/Q17 M) Neor ′amyE | This work |
| pCG2 | bla amyE′ PspaS BamHI Neor ′amyE | This work |
| pTSp99 | bla amyE′ PspaS-spaS(W1E) Neor ′amyE | This work |
| pTSp100 | bla amyE′ PspaS-spaS(W1Y) Neor ′amyE | This work |
| pTSp101 | bla amyE′ PspaS-spaS(W1F) Neor ′amyE | This work |
| pTSp102 | bla amyE′ PspaS-spaS(W1G) Neor ′amyE | This work |
| pTSp108 | bla amyE′ PspaS-spaS(W1D) Neor ′amyE | This work |
| pTSp109 | bla amyE′ PspaS-spaS(W1A) Neor ′amyE | This work |
| pTSp110 | bla amyE′ PspaS-spaS(K2T/E4I/V12K/Q17M) Neor ′amyE | This work |
| pTSp112 | bla amyE′ PspaS-spaS(W1H) Neor ′amyE | This work |
| pTSp115 | bla amyE′ PspaS-spaS(W1K) Neor ′amyE | This work |
| pTSp116 | bla amyE′ PspaS-spaS(W1R) Neor ′amyE | This work |
| pTSp124 | bla amyE′ PspaS-spaS(W1L) Neor ′amyE | This work |
| pTSp125 | bla amyE′ PspaS-spaS(W1V) Neor ′amyE | This work |
| pNZ8148 | Vector carrying nisA promoter Cmr | 38 |
| pSB5 | bla amyE′ PspaS-lacZ Cmr ′amyE | 18 |
| pAUT3 | pNZ8148 PnisA-lacZ | This work |
a
Ent+, entianin producer; Ent−, entianin nonproducer; Sub+, subtilin producer; Sub−, subtilin nonproducer; Cmr, chloramphenicol resistant; Specr, spectinomycin resistant; Neor, neomycin resistant. Letters in parentheses indicate the amino acid residues exchanged (corresponding to the IUPAC abbreviation), and numbers in parentheses indicate the positions of the exchanged amino acids in the respective subtilin variants. BGSC, Bacillus Genetic Stock Center.
b
DSM, German Resource Centre for Biological Material; ATCC, American Type Culture Collection.