Table 3.
Strains and plasmids used in this study
| Strain/plasmid | Characteristic | Source |
|---|---|---|
| Strain | ||
| E. coli DH5ɑ | Clone host strain | Transgen |
| Orn8 | C. glutamicum S9114 with deletion of argF, ncgl1221, argR, putP, attenuation of odhA, and overexpression of lysE | Lab stock |
| Orn17 | Orn8 carrying expression vector pEC-XK99E | This study |
| Orn18 | Orn8 carrying expression vector pEC-CgS09558 | This study |
| Orn19 | Orn8 carrying expression vector pEC-CgS14252 | This study |
| Orn20 | Orn8 with Ptac promoter inserted in front of CgS9114_09558 | This study |
| Orn21 | Orn8 with Ptac promoter inserted in front of CgS9114_14252 | This study |
| Orn22 | Orn8 with deletion of CgS9114_13845 | This study |
| Orn23 | Orn8 with deletion of CgS9114_02593 | This study |
| Orn24 | Orn8 with deletion of CgS9114_02058 | This study |
| Orn25 | Orn8 with deletion of CgS9114_14352 | This study |
| Orn26 | Orn8 with deletion of CgS9114_04952 | This study |
| Orn27 | Orn8 with deletion of CgS9114_14497 | This study |
| Orn28 | Orn8 with deletion of CgS9114_13845 and CgS9114_02593 | This study |
| Orn29 | Orn8 with deletion of CgS9114_13845, CgS9114_02593 and CgS9114_02058 | This study |
| Orn30 | Orn8 with deletion of CgS9114_13845, CgS9114_02593, and CgS9114_02058; overexpression of CgS9114_09558 | This study |
| Plasmid | ||
| pK18mobsacB | Mobilizable vector, allows for selection of double crossover in C. glutamicum, KmR, sacB | [16] |
| pEC-XK99E | A shuttle expression vector, KmR | Lab stock |
| pEC-CgS14252 | A derivative of pEC-XK99E, harboring CgS9114_14252 gene from C. glutamicum S9114 under Peftu promoter | This study |
| pEC-CgS09558 | A derivative of pEC-XK99E, harboring CgS9114_09558 gene from C. glutamicum S9114 under Peftu promoter | This study |
| pK18-Ptac-CgS09558 | A derivative of pK18mobsacB, harboring Ptac-CgS09558 fragment | This study |
| pK18-Ptac-CgS14252 | A derivative of pK18mobsacB, harboring Ptac-CgS14252 fragment | This study |
| pK18-ΔCgS13845 | A derivative of pK18mobsacB, harboring ΔCgS9114_13845 fragment | This study |
| pK18-ΔCgS02593 | A derivative of pK18mobsacB, harboring ΔCgS9114_02593 fragment | This study |
| pK18-ΔCgS02058 | A derivative of pK18mobsacB, harboring ΔCgS9114_02058 fragment | This study |
| pK18-ΔCgS14352 | A derivative of pK18mobsacB, harboring ΔCgS9114_14352 fragment | This study |
| pK18-ΔCgS04952 | A derivative of pK18mobsacB, harboring ΔCgS9114_04952 fragment | This study |
| pK18-ΔCgS14497 | A derivative of pK18mobsacB, harboring ΔCgS9114_14497 fragment | This study |
Superscript ‘‘R’’ indicates resistance to the following antibiotics: Km kanamycin