Table 1 Strains and plasmids used in this study
Strain/plasmid | Characteristic | Source |
|---|---|---|
Strain | ||
E. coli DH5ɑ | Clone host strain | Transgen |
SO1 | C. glutamicum S9114 with deletion of argF, ncgl1221, argR, putP, attenuation of odhA, proB, ncgl2228, overexpression of lysE, gdh, argCJBD | Lab stock |
SO2 | SO1 carrying expression vector pEC-XK99E | This study |
SO3 | SO1 carrying expression vector pEC-cg3035 | This study |
SO4 | SO1 with P tac promoter inserted in front of cg3035 | This study |
SO5 | SO1 with terminator inserted in front of pta | This study |
SO6 | SO1 with terminator inserted in front of cat | This study |
SO7 | SO1 with P eftu promoter inserted in front of pfkA | This study |
SO8 | SO1 with P eftu promoter inserted in front of gap | This study |
SO9 | SO1 with P eftu promoter inserted in front of pyk | This study |
SO10 | SO1 with P eftu promoter inserted in front of pfkA and P tac promoter inserted in front of cg3035 | This study |
SO11 | SO1 with P eftu promoter inserted in front of pfkA, P tac promoter inserted in front of cg3035 and a terminator inserted in front of pta | This study |
SO12 | SO1 with P eftu promoter inserted in front of pfkA, P tac promoter inserted in front of cg3035 and a terminator inserted in front of cat | This study |
SO13 | SO1 with P eftu promoter inserted in front of pfkA, P tac promoter inserted in front of cg3035 and terminator inserted in front of pta and cat | This study |
SO14 | SO12 with an AAV degradation label inserted in the C terminal of pgi | This study |
SO15 | SO14 with synthetic RBS and A1G replacement in front of zwf. | This study |
SO16 | SO15 with P tac promoter inserted in front of tkt operon. | 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-cg3035 | A derivative of pEC-XK99E, harboring cg3035 gene from C. glutamicum S9114 under its native promoter | This study |
pK18-P tac -cg3035 | A derivative of pK18mobsacB, harboring P tac -cg3035 fragment | This study |
pK18-T-pta | A derivative of pK18mobsacB, harboring T-pta fragment | This study |
pK18- T-cat | A derivative of pK18mobsacB, harboring T-cat fragment | This study |
pK18-P eftu -pfkA | A derivative of pK18mobsacB, harboring P eftu -pfkA fragment | This study |
pK18-P eftu -gap | A derivative of pK18mobsacB, harboring P eftu -gap fragment | This study |
pK18-P eftu -pyk | A derivative of pK18mobsacB, harboring P eftu -pyk fragment | This study |
pK18- pgi-aav | A derivative of pK18mobsacB, harboring pgi-AAV fragment | This study |
pK18-SB-zwf | A derivative of pK18mobsacB, harboring SB-zwf fragment | This study |
pK18-P tac -tkt | A derivative of pK18mobsacB, harboring P tac -tkt fragment | This study |