TABLE 1.
Plasmids and bacterial strains used in this study
| Strain/plasmid | Relevant genotypea | Reference |
|---|---|---|
| S. pyogenes | ||
| NV1 | GAS 5448 serotype M1T1 | (12) |
| NV2 | NV1, cat, pAV488 (cat, recombineering plasmid) | This study |
| NV3 | NV2, cat, ery, pAV488, ΔhsdR::ery | This study |
| NV4 | NV3, cat, ery, kan, pAV488, ΔhsdR::ery, Δcas9::kan | This study |
| NV6 | NV1, ery, tet, ΔhsdR::ery, Δcas9::Ptet-dcas9, tetR, tetM | This study |
| NV9 | NV1, cat, pDC-sgRNA (P3-mCherry-sgRNA, cat) | This study |
| NV19 | NV6, ery, tet, cat, ΔhsdR::ery, Δcas9::Ptet-dcas9, tetR, tetM, pDC-sgRNA-ftsZ (P3-sgRNA-ftsZ, cat) | This study |
| NV22 | NV6, ery, tet, cat, ΔhsdR::ery, Δcas9::Ptet-dcas9, tetR, tetM, pDC-sgRNA-dnaA (P3-sgRNA-dnaA, cat) | This study |
| NV25 | NV6, ery, tet, cat, ΔhsdR::ery, Δcas9::Ptet-dcas9, tetR, tetM, pDC-sgRNA-emm (P3-sgRNA-emm, cat) | This study |
| NV26 | NV6, ery, tet, cat, ΔhsdR::ery, Δcas9::Ptet-dcas9, tetR, tetM, pDC-sgRNA-control (P3-sgRNA-control, cat) | This study |
| Escherichia coli | ||
| MC1061 | F- Δ(ara-leu)7697 [araD139]B/r Δ(codB-lacI)3 galK16 galE15 λ- e14- mcrA0 relA1 rpsL150(StrR) spoT1 mcrB1 hsdR2(r-m+) | MClab |
| Plasmids | ||
| pDC123 | cat, phoZ | (24) |
| pDC-sgRNA | P3-mCherry-sgRNA, cat | This study |
a
cat, chloramphenicol resistance marker; ery, erythromycin resistance marker; tetM, tetracycline resistance marker; and kan, kanamycin resistance marker.