TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant characteristics or sequence (5′ to 3′)a | Source or purpose |
|---|---|---|
| Strains | ||
| E. coli DH5α | F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | 20 |
| K. europaeus | ||
| KGMA0119 | Wild type isolated from rice vinegar | 18 |
| KGMA0704 | KGMA0119 derivative; ΔpyrE | 18 |
| KGMA5511 | KGMA0704 derivative; ΔpyrE ΔilvC::pyrE | This work |
| KGMA7203 | KGMA0704 derivative; ΔpyrE KelrpΔ(397-511)::pyrE | 19 |
| Plasmids | ||
| pBR322 | Ampr | 21 |
| pBR322-ΔilvC::pyrE | pBR322 derivative; ΔilvC::pyrE cassette composed of 5′ upstream and 3′ downstream regions of ilvC and a pyrE cassette (3,374 bp) | This work |
| Primers | ||
| ilvC-F | AAGAATTCTCACGGTATTCCGAAAGCTACAGT | ilvC disruption |
| ilvC-R | AAGGATCCGTATTCAGGCGGTATAGGTGACGTT | ilvC disruption |
| ilvC-i1 | ATGATGTCATGGATCGGCAAGAACAAGCTG | ilvC disruption |
| ilvC-i2 | ATCGCGATCGTAATAGACGCGCATGGTCTT | ilvC disruption |
| EP-F2 | CTGCCATATCCCGTGTTCGT | ilvC disruption |
| E-R4 | TCGCCATAGGGAAAGACTGC | ilvC disruption |
| ilvC-F2 | GTGGATACAATATCGAAAGCCTGAC | Genotyping |
| ilvC-R2 | CTGGCTTGGAAAAGAAGAAAGGTTC | Genotyping |
a
Restriction enzyme sites are underlined. Amp, ampicillin.