Table 1.
Bacterial strains and plasmids used in this study.
| Genotype | Source or reference | |
|---|---|---|
| E. coli STRAIN | ||
| BL21(DE3) | Expressing host cell | Laboratory collection |
| BL21(DE3) pET30a::cas1 | E. coli BL21(DE3) carrying pET30a::cas1, KanR | This study |
| BL21(DE3) pET30a::cas1E149A | E. coli BL21(DE3) carrying pET30a::cas1E149A, KanR | This study |
| BL21(DE3) pET30a::cas1H206A | E. coli BL21(DE3) carrying pET30a::cas1H206A, KanR | This study |
| BL21(DE3) pET30a::cas1E221A | E. coli BL21(DE3) carrying pET30a::cas1E221A, KanR | This study |
| BL21(DE3) pET30a::cas1-cas2 | E. coli BL21(DE3) carrying pET30a::cas1-cas2, KanR | This study |
| DH5a pGEM::CRISPR | E. coli DH5a carrying pGEM::CRISPR, AmpR | This study |
| R. anatipestifer STRAIN | ||
| RA-CH-2 | Serotype 2, CfxS, SpcS | Laboratory collection |
| RA-CH-2 pLMF03 | RA-CH-2 carrying pLMF03, CfxR, SpcS | This study |
| RA-CH-2 Δcas1 | cas1 deletion mutant of R. anatipestifer CH-2 strain, CfxS, SpcR | This study |
| RA-CH-2 Δcas2 | cas2 deletion mutant of R. anatipestifer CH-2 strain, CfxS, SpcR | This study |
| RA-CH-2 Δcas1-cas2 | cas1-cas2 deletion mutant of R. anatipestifer CH-2 strain, CfxS, SpcR | This study |
| RA-CH-2 Δcas1 pLMF03 | RA-CH-2 Δcas1 carrying pLMF03, CfxR, SpcR | This study |
| RA-CH-2 Δcas2 pLMF03 | RA-CH-2 Δcas2 carrying pLMF03, CfxR, SpcR | This study |
| RA-CH-2 Δcas1-cas2 pLMF03 | RA-CH-2 Δcas1-cas2 carrying pLMF03, CfxR, SpcR | This study |
| PLASMID | ||
| pET30a | pBR322 lacZ, IPTG-inducible promoter, KanR | Laboratory collection |
| pET30a::cas1 | pET30a carrying cas1 from R. anatipestifer CH-2 adding His tag, KanR | This study |
| pET30a::cas1E149A | pET30a carrying cas1E149A adding His tag, KanR | This study |
| pET30a::cas1H206A | pET30a carrying cas1H206A adding His tag, KanR | This study |
| pET30a::cas1E221A | pET30a carrying cas1E221A adding His tag, KanR | This study |
| pET30a::cas1-cas2 | pET30a carrying cas1-cas2 from R. anatipestifer CH-2 adding His tag and Flag tag, KanR | This study |
| pLMF03 | B739_0921 promoter, ori ColE1, ori pRA0726, AmpR, CfxR | Liu M. et al., 2016 |