Table 3.
Gene editing in this study
| Strains and Genes | Locus | Plasmids | N20 and PAM sites | LA (bp)a |
|---|---|---|---|---|
|
Gene deletion C. glutamicum ATCC 13032 | ||||
| ldh | NCgl2810 | PCCG1-ldh | CAGCATGTAGCGGAATCGAGCGG | 916/989 |
| eutD | NCgl2657 | pCCG1-eutD | GTCGCCGTTGTGGACCATCATGG | 800/839 |
| gabP | NCgl0464 | pCCG1-gabP | GTGGAGGCATTGATCACCACTGG | 553/580 |
| glnA1 | NCgl2133 | pCCG1-glnA1 | CGTGGAGCATGGTGTTGAAGCGG | 700/643 |
| glnA2 | NCgl2148 | pCCG1-glnA2 | GAAATATCCGCCGTTGTCAGTGG | 609/630 |
| alaT | NCgl2747 | pCCG1-alaT | GGTTGCCAGGCTGATGTGCTCGG | 739/765 |
| argR | NCgl1345 | pCCG1-argR | TCGCGGGCGATGCTATCTACTGG | 564/625 |
| gabTD | NCgl0462, NCgl0463 | pCCG1-gabTD | CCAGTAATCAACCCCAGCGATGG | 754/754 |
| aceAB | NCgl2248, NCgl2247 | pCCG1-aceAB | GGAAATCCTCGTACGCCTCTTGG | 810/829 |
| poxB | NCgl2521 | pCCG1-poxB | GGTCACCGGATACTTCACCGTGG | 644/654 |
| C. glutamicum ATCC 14067 | ||||
| gabP | CEY17_02745 | pCCG1-gabP-2 | GTGGAGGCATTGATCACCACTGG | 553/580 |
| C. glutamicum ssp. lactofermentum | ||||
| gabP | NDb | pCCG1-gabP-3 | GTGGAGGCATTGATCACCACTGG | 553/580 |
| Gene insertion | ||||
| Φ(PtacM-gdhA1)c | NCgl1999 | pCCG1-PtacM-gdhA1 | GCAAACGCCTAGGATGTACATGG | 721/662 |
| eutD::speC | NCgl2657 | pCCG1-ΔeutD::speC | GTCGCCGTTGTGGACCATCATGG | 800/839 |
| gabP::gadB | NCgl0464 | pCCG1-ΔgabP::gadB | GTGGAGGCATTGATCACCACTGG | 553/580 |
| gabP::gadB2B1m | NCgl0464 | pCCG1-ΔgabP::gadB2-gadB1m | GTGGAGGCATTGATCACCACTGG | 553/580 |
| eutD::gdhA | NCgl2657 | pCCG1-ΔeutD::gdhA | GTCGCCGTTGTGGACCATCATGG | 800/849 |
| alaT::gdhA | NCgl2747 | pCCG1-ΔalaT::gdhA | GGTTGCCAGGCTGATGTGCTCGG | 739/765 |
| alaT::gdhA | NCgl2747 | pCCG2-ΔalaT::gdhA | GGTTGCCAGGCTGATGTGCTCGG | 739/765 |
a, LA: the length of homologous arms (bp), which is the up and down homologous sequence flanking the deletion target sequence; b, ND: no data; c, Using the strong promoter PtacM and artificial RBS (GAAAGGACTTGAACG) sequence to replace the native promoter of gene gdhA1 in genome