TABLE 1.
C. glutamicum strains, relevant plasmids, and plasmids used for overexpression together with relevant primers
| Strain designation | Description or primer sequencea |
|---|---|
| Strains | |
| ATCC 13032 | Wild type (WT) |
| WTΔodhA | Chromosomal deletion of nt 1174001-1177720 |
| WTΔodhAE1 | Chromosomal deletion of nt 1174001-1176601 |
| WTΔodhAE2 | Chromosomal deletion of nt 1176641-1177330 |
| WTΔodhAE1L | Chromosomal deletion of nt 1177361-1177624 |
| WTΔaceF | Chromosomal deletion of nt 2310962-2312939 |
| WTΔaceE | Chromosomal deletion of aceE, obtained from reference 31 |
| WT aceF(K43Q-K162Q) | Chromosomal point mutations to result in AceF-K43Q-K162Q |
| WT aceF(K162Q-K278Q) | Chromosomal point mutations to result in AceF-K162Q-K278Q |
| Plasmids | |
| pK19mobsacB-K43Q-K162Q | pK19mobsacB containing custom-synthesized 1,240-bp fragment of aceF |
| pK19mobsacB-K162Q-K278Q | pK19mobsacB containing custom-synthesized 1,200-bp fragment of aceF |
| pEKEx5-H6-AceE | AATGGCCGATCAAGCAAAACTTG |
| TTATTCCTCAGGAGCGTTTGGAT | |
| pEKEx5-H6-AceF | CGCGGATCCGCGATGGCGTTCTCCGTAGAGATGC |
| CGCGGATCCGCGGAGCTGCAGATCGCCTTCGAAG | |
| pEKEx2-odh′-H6 | CTGAGCTAGGATCCCTCTACGACCCCAATGAAGG |
| TAGCTCAGGAATTCATTGTGATGGTGATGGTGATGGGCAGCCTTTGGTGCTGGTT | |
| pEKEx5-odhA-T294A | CGTTACCGTTTCCTTGGCCAACCCAGGTGGCATC |
| GATGCCACCTGGGTTGGCCAAGGAAACGGTAACG | |
| pEKEx5-odhA-H352C | CACCTCCACCTACGATTGCCGCGTGATCCAGGGTG |
| CACCCTGGATCACGCGGCAATCGTAGGTGGAGGTG | |
| pEKEx5-odhA-Q356D | CTACGATCACCGCGTGATCGATGGTGCTGTGTCCGGTG |
| CACCGGACACAGCACCATCGATCACGCGGTGATCGTAG | |
| pEKEx5-odhA-H352C-Q-356D | CTCCACCTACGATTGCCGCGTGATCGATGGTGCTGTGTCCGGTGA |
| TCACCGGACACAGCACCATCGATCACGCGGCAATCGTAGGTGGAG |
a
The nucleotide positions refer to the genome sequence of NC_006958.1. Primer sequences are given in direction 5′ to 3′. The primers given for the pEKEx5 plasmids show those with mutations in odhA with the mutated nucleotides in italics. nt, nucleotides.