Table 2.
Bacterial strains and plasmids used in this study
| Relevant characteristics | Reference or source | |
|---|---|---|
| Strain | ||
| E. coli | ||
| XL1-blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proABlacIq ZΔM15 Tn10 (Tet r )] | Stratagenea |
| C. glutamicum | ||
| ATCC 13032 | Biotin-auxotrophic wild type | ATCC |
| WJ001 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg1a) | This study |
| WJ002 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg1a, ISCg1b) | This study |
| WJ003 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg1a, ISCg1b, ISCg1c) | This study |
| WJ004 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg1a, ISCg1b, ISCg1c, ISCg1e) | This study |
| WJ005 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg2b) | This study |
| WJ006 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg2b, ISCg2c) | This study |
| WJ007 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg2b, ISCg2c, ISCg2e) | This study |
| WJ008 | ATCC 13032 with in frame deletion of ISCg2 family (ISCg2b, ISCg2c, ISCg2e, ISCg2f) | This study |
| Plasmids | ||
| pCES208 | E. coli—C. glutamicum shuttle vector, Kmr | [45] |
| pCES-NMCS | pCES208 derivative; MCS and rrn terminator, Kmr | [6] |
| pH36M2 | pCES208 derivative; PH36, PorB signal sequence, codon-optimized M18 scFv gene (opt) | [5] |
| pCES-H36-porBss | pCES208 derivative; PH36, porinB signal sequence, Kmr | This study |
| pCES-H36-porBss-Amy | pCES208 derivative; PH36, porinB signal sequence amylase, Kmr | This study |
| pCES-H36-porBss-IS-Amy | pCES208 derivative; PH36, IS30 inserted porinB signal sequence, amylase, Kmr | This study |
| pCES-H36-GFP | pCES208 derivative; PH36, eGFP, Kmr | [46] |
| pCnCAB | phaCABC gene expression | [41] |
| pCES-H36-PhaC | pCES208 derivative; PH36, phaC, Kmr | This study |
| pCES-H36-PhaCA | pCES208 derivative; PH36, phaCA, Kmr | This study |
| pCES-H36-PhaCAB | pCES208 derivative; PH36, phaCAB, Kmr | This study |
| pHGmut | pCES208 derivative, Glu89Gln/Δ452-466 gene | [26] |
| pK19mobsacB | Mobilizable vector, Kmr | [42] |
| pK19-∆ISCg1a | pK19mobsacB derivative; flanking region of ISCg1a | This study |
| pK19-∆ISCg1b | pK19mobsacB derivative; flanking region of ISCg1b | This study |
| pK19-∆ISCg1c | pK19mobsacB derivative; flanking region of ISCg1c | This study |
| pK19-∆ISCg1e | pK19mobsacB derivative; flanking region of ISCg1e | This study |
| pK19-∆ISCg2b | pK19mobsacB derivative; flanking region of ISCg2b | This study |
| pK19-∆ISCg2c | pK19mobsacB derivative; flanking region of ISCg2c | This study |
| pK19-∆ISCg2e | pK19mobsacB derivative; flanking region of ISCg2e | This study |
| pK19-∆ISCg2f | pK19mobsacB derivative; flanking region of ISCg2f | This study |
| pJET1.2/blunt | Commercial cloning vector | Thermob |
aNew England Biolabs, Beverly, MA, USA
bThermo fisher scientific, Waltham, MA, USA