Table 1.
Strains and plasmids used in this study.
| Plasmids/strains | Description | Sources |
|---|---|---|
| Plasmids | ||
| pCM80 | vector used for gene expression in M. extorquens; promoter, PmxaF; antibiotics, TetR, KmR | [28] |
| pCM433 | sacB-based allelic exchange vector; antibiotics, ApR, CmR, TetR | [29] |
| pAIO | pCM80 derivative with dcas9 and sgRNA expression cassette; dcas9 and sgRNA under the control of inducible promoter PR/tetO and PmxaF-g, respectively | [26] |
| pAI1815 | pAIO derivative plasmid carrying the sgRNA targeting META1p1815 | This study |
| pAI1816 | pAIO derivative plasmid carrying the sgRNA targeting META1p1816 | This study |
| pCM3663 | pCM80 derivative harboring META1p3663 under the downstream of PmxaF, TetR | This study |
| pCM3664 | pCM80 derivative harboring META1p3664 under the downstream of PmxaF, TetR | This study |
| pCM1815 | pCM80 derivative harboring META1p1815 under the downstream of PmxaF, TetR | This study |
| pCM1816 | pCM80 derivative harboring META1p1816 under the downstream of PmxaF, TetR | This study |
| pCM1815-16 | pCM80 derivative harboring the operon of META1p1815 and META1p1816 under the downstream of PmxaF, TetR | This study |
| pCM3665-70 | pCM80 derivative harboring the operon of META1p3665 and META1p3670 under the downstream of PmxaF, TetR | This study |
| pCM1815-3665-70 | pCM80 derivative harboring the operon of META1p1815, META1p3665 and META1p3670 under the downstream of PmxaF, TetR | This study |
| pCM-△3663 | pCM433 with about 1 kb upstream and 1 kb downstream fragments of META1p3663; ApR, CmR, TetR | This study |
| pCM-△3664 | pCM433 with about 1 kb upstream and 1 kb downstream fragments of META1p3664; ApR, CmR, TetR | This study |
| pCM-△3652 | pCM433 with about 1 kb upstream and 1 kb downstream fragments of META1p3652; ApR, CmR, TetR | This study |
| pCM-△4598 | pCM433 with about 1 kb upstream and 1 kb downstream fragments of META1p4598; ApR, CmR, TetR | This study |
| Strains | ||
| E. coli DH5αα | Host strain for general clone | Lab storage |
| M. extorquens AM1 | Wild-type, pink color, rifamycin-resistant strain | [30] |
| YA | M. extorquens AM1::pCM80 | [26] |
| YAIO | M. extorquens AM1 containing the plasmid pAIO | [26] |
| YA3663 | The gene META1p3663 deleted from M. extorquens AM1 | This study |
| YA3664 | The gene META1p3664 deleted from M. extorquens AM1 | This study |
| YA3652 | The gene META1p3652 deleted from M. extorquens AM1 | This study |
| YA4598 | The gene META1p4598 deleted from M. extorquens AM1 | This study |
| YCM3663 | The strain YA3663::pCM-exp3663, TetR | This study |
| YCM3664 | The strain YA3664::pCM-exp3664, TetR | This study |
| YAMZ1815 | M. extorquens AM1 harboring the CRISPRi plasmid pAI1815 | This study |
| YAMZ1816 | M. extorquens AM1 harboring the CRISPRi plasmid pAI1816 | This study |
| YCM1815 | M. extorquens AM1::pCM-exp1815, TetR | This study |
| YCM1816 | M. extorquens AM1::pCM-exp1816, TetR | This study |
| YCM1815-16 | M. extorquens AM1::pCM-exp1815-16, TetR | This study |
| YCM3665-70 | M. extorquens AM1::pCM-exp3665-70, TetR | This study |
| YCM1816-3665-70 | M. extorquens AM1:: pCM-exp1815-3665-70, TetR | This study |