Table 2.
Strains and plasmids used in this study.
| Name | Description | Source |
|---|---|---|
| Strains | ||
| C. glutamicum S9114 ΔnagA-ΔgamA-Δldh | C. glutamicum S9114 derivate: ΔnagA-ΔgamA-Δldh | Laboratory stock |
| E. coli JM109 | Laboratory stock | |
| M1 | C. glutamicum S9114 ΔnagA-ΔgamA-Δldh embedding zwf mutant P286L | This work |
| M2 | C. glutamicum S9114 ΔnagA-ΔgamA-Δldh embedding zwf mutants P286L, A287G | This work |
| M3 | C. glutamicum S9114 ΔnagA-ΔgamA-Δldh embedding zwf mutant P286S | This work |
| M4 | C. glutamicum S9114 ΔnagA-ΔgamA-Δldh embedding zwf mutant L290stop | This work |
| M5 | C. glutamicum S9114 ΔnagA-ΔgamA-Δldh embedding zwf mutants P286L, K293E | This work |
| M6 | C. glutamicum S9114 ΔnagA-ΔgamA-Δldh embedding zwf mutant K293R | This work |
| Plasmids | ||
| pJYW-4-ceN | pJYW-4 derivate, Ptac-GNA1,Kanr | Laboratory stock |
| pXMJ19 | Cloning vector, Cmr | Laboratory stock |
| pXMJ19-Cas9-AID pXMJ19-dCas9-AID |
pXMJ19 derivate with Cas9 and AID cloned pXMJ19-Cas9-AID with D10A and H840A mutation sites |
This work This work |
| pFST-porb | pFST with sgRNA expression framework cloned | Laboratory stock |
| pXMJ19-dCas9 1 | pXMJ19-dCas9-AID with sgRNA expression framework cloned | This work |
| pXMJ19-dCas9 1-zwf1 | pXMJ19-dCas9 1 with PAM1 of gene zwf | This work |
| pXMJ19-dCas9 1-zwf2 | pXMJ19-dCas9 1 with PAM2 of gene zwf | This work |
| pXMJ19-dCas9 1-zwf3 | pXMJ19-dCas9 1 with PAM3 of gene zwf | This work |
| pXMJ19-dCas9 1-zwf4 | pXMJ19-dCas9 1 with PAM4 of gene zwf | This work |
| pMD19-T | Laboratory stock | |
| pMD19-T Vector23100 | pMD19-T derivate with sgRNA framework gene under the control of Pj23100 | This work |
| pMD19-T Vector23102 | pMD19-T derivate with sgRNA framework gene under the control of Pj23102 | This work |
| pMD19-T Vector23104 | pMD19-T derivate with sgRNA framework gene under the control of Pj23104 | This work |
| pMD19-T Vector23105 | pMD19-T derivate with sgRNA framework gene under the control of Pj23105 | This work |
| pMD19-T Vector23108 | pMD19-T derivate with sgRNA framework gene under the control of Pj23108 | This work |
| pMD19-T Vector23111 | pMD19-T derivate with sgRNA framework gene under the control of Pj23111 | This work |
| pMD19-T Vector23113 | pMD19-T derivate with sgRNA framework gene under the control of Pj23113 | This work |
| pMD19-T Vector23118 | pMD19-T derivate with sgRNA framework gene under the control of Pj23118 | This work |
| pMD19-T Vector23119 | pMD19-T derivate with sgRNA framework gene under the control of Pj23119 | This work |
| pXMJ19-dCas9 2-23100 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23100 | This work |
| pXMJ19-dCas9 2-23102 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23102 | This work |
| pXMJ19-dCas9 2-23104 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23104 | This work |
| pXMJ19-dCas9 2-23105 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23105 | This work |
| pXMJ19-dCas9 2-23108 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23108 | This work |
| pXMJ19-dCas9 2-23111 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23111 | This work |
| pXMJ19-dCas9 2-23113 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23113 | This work |
| pXMJ19-dCas9 2-23118 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23118 | This work |
| pXMJ19-dCas9 2-23119 | pXMJ19-dCas9 1 derivate with sgRNA framework gene under the control of Pj23119 | This work |
| pXMJ19-dCas9 2-23100-P | pXMJ19-dCas9 2-23100 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23100-P | pXMJ19-dCas9 2-23100 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23102-P | pXMJ19-dCas9 2-23102 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23104-P | pXMJ19-dCas9 2-23104 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23105-P | pXMJ19-dCas9 2-23105 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23108-P | pXMJ19-dCas9 2-23108 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23111-P | pXMJ19-dCas9 2-23111 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23113-P | pXMJ19-dCas9 2-23113 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23118-P | pXMJ19-dCas9 2-23118 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 2-23119-P | pXMJ19-dCas9 2-23119 derivate with 20N sequence and PAM site AGG | This work |
| pXMJ19-dCas9 3 | pXMJ19-dCas9 2-23119-P derivate with TadA-TadA mutant cloned | This work |
| pTYW-4-ceN-C.glglmS | pJYW-4-ceN derivate with C.glglmS cloned, Kanr | Laboratory stock |
Cmr, chloramphenicol resistance; Kanr, kanamycin resistance.