TABLE 3.
Plasmid constructs used in the present study
| Name | Description | Restriction site(s)a | Source or reference |
|---|---|---|---|
| pCRB214 | Kmr; cloning vector with Ptac and rrnBT1T2 derived from pCASE1 | NA | 44 |
| pCASE1 | Plasmid identified in Corynebacterium casei JCM12072 | NA | 69 |
| pCRB12 | Kmr; pCG1-based C. glutamicum-E. coli shuttle vector | NA | 43 |
| pCG1 | Plasmid identified in C. glutamicum ATCC 31808 | NA | 70 |
| pDW363 | Apr; source of lacIq | NA | NBRP (NIG, Japan) |
| pCRB12i | Kmr; IPTG-inducible vector derived from pCRB12 | NA | This study |
| pHSG298 | Kmr; α-lac multicloning site; E. coli cloning vector | NA | TaKaRa Bio, Inc. |
| pCRA725 | Kmr; pHSG298 with Ptac-sacR-sacB | NA | 45 |
| pCRE500 | GLY3 ntb 39536–41166 in pCRA725; plasmid for allelic replacement of CgR_0035 | XbaI/SalI | This study |
| pCRE501 | tam45 nt 39536–41166 in pCRA725; plasmid for allelic replacement of CgR_0035 | XbaI/SalI | This study |
| pCRE502 | GLY3 nt 157063–158683 in pCRA725; plasmid for allelic replacement of CgR_0141 | SalI | This study |
| pCRE503 | tam45 nt 157063–158683 in pCRA725; plasmid for allelic replacement of CgR_0141 | SalI | This study |
| pCRE504 | GLY3 nt 545101–546040 in pCRA725; plasmid for allelic replacement of hemC | XbaI/SalI | This study |
| pCRE505 | tam45 nt 545101–546040 in pCRA725; plasmid for allelic replacement of hemC | XbaI/SalI | This study |
| pCRE506 | GLY3 nt 1151531–1153602 in pCRA725; plasmid for allelic replacement of glmU | XbaI/SalI | This study |
| pCRE507 | tam45 nt 1151531–1153602 in pCRA725; plasmid for allelic replacement of glmU | XbaI/SalI | This study |
| pCRE508 | GLY3 nt 1179149–1180226 in pCRA725; plasmid for allelic replacement of CgR_1064 | SalI | This study |
| pCRE509 | tam45 nt 1179149–1180226 in pCRA725; plasmid for allelic replacement of CgR_1064 | SalI | This study |
| pCRE510 | GLY3 nt 1356603–1357635 in pCRA725; plasmid for allelic replacement of mhpA | SalI | This study |
| pCRE511 | tam45 nt 1356603–1357635 in pCRA725; plasmid for allelic replacement of mhpA | SalI | This study |
| pCRE512 | GLY3 nt 1423170–1424283 in pCRA725; plasmid for allelic replacement of nucS | SalI | This study |
| pCRE513 | tam45 nt 1423170–1424283 in pCRA725; plasmid for allelic replacement of nucS | SalI | This study |
| pCRE514 | GLY3 nt 1899141–1900182 in pCRA725; plasmid for allelic replacement of secN | SalI | This study |
| pCRE515 | tam45 nt 1899141–1900182 in pCRA725; plasmid for allelic replacement of secN | SalI | This study |
| pCRE516 | trm2 nt 1899141–1900182 in pCRA725; plasmid for allelic replacement of secN | SalI | This study |
| pCRE517 | GLY3 nt 2257572–2259106 in pCRA725; plasmid for allelic replacement of mraW | XbaI/SalI | This study |
| pCRE518 | tam45 nt 2257572–2259106 in pCRA725; plasmid for allelic replacement of mraW | XbaI/SalI | This study |
| pCRE519 | GLY3 nt 2267254–2268257 in pCRA725; plasmid for allelic replacement of pknL | SalI | This study |
| pCRE520 | tam45 nt 2267254–2268257 in pCRA725; plasmid for allelic replacement of pknL | SalI | This study |
| pCRE521 | GLY3 nt 2385305–2386847 in pCRA725; plasmid for allelic replacement of hrcA | SalI | This study |
| pCRE522 | trm2 nt 2385305–2386847 in pCRA725; plasmid for allelic replacement of hrcA | SalI | This study |
| pCRE523 | tam45 nt 2385305–2386847 in pCRA725; plasmid for allelic replacement of hrcA | SalI | This study |
| pCRE524 | GLY3 nt 2537845–2538846 in pCRA725; plasmid for allelic replacement of CgR_2303 | SalI | This study |
| pCRE525 | tam45 nt 2537845–2538846 in pCRA725; plasmid for allelic replacement of CgR_2303 | SalI | This study |
| pCRE526 | GLY3 nt 2566914–2567767 in pCRA725; plasmid for allelic replacement of CgR_2333 | XbaI/SalI | This study |
| pCRE527 | tam45 nt 2566914–2567767 in pCRA725; plasmid for allelic replacement of CgR_2333 | XbaI/SalI | This study |
| pCRE528 | GLY3 nt 2797092–2798776 in pCRA725; plasmid for allelic replacement of otsA | SacI/SalI | This study |
| pCRE529 | tam45 nt 2797092–2798776 in pCRA725; plasmid for allelic replacement of otsA | SacI/SalI | This study |
| pCRE530 | GLY3 nt 2812261–2814354 in pCRA725; plasmid for allelic replacement of ptsS | XbaI/SalI | This study |
| pCRE531 | trm2 nt 2812261–2814354 in pCRA725; plasmid for allelic replacement of ptsS | XbaI/SalI | This study |
| pCRE532 | tam45 nt 2812261–2814354 in pCRA725; plasmid for allelic replacement of ptsS | XbaI/SalI | This study |
| pCRE533 | GLY3 nt 2936034–2937632 in pCRA725; plasmid for allelic replacement of pknG | XbaI | This study |
| pCRE534 | tam45 nt 2936034–2937632 in pCRA725; plasmid for allelic replacement of pknG | XbaI | This study |
| pCRE535 | GLY3 nt 3297021–3298018 in pCRA725; plasmid for allelic replacement of CgR_2975 | XbaI/SalI | This study |
| pCRE536 | tam45 nt 3297021–3298018 in pCRA725; plasmid for allelic replacement of CgR_2975 | XbaI/SalI | This study |
| pCRE537 | GLY3 nt 1122184–1123706 in pCRA725; plasmid for allelic replacement of CgR_1015-1016 | XbaI/SalI | This study |
| pCRE538 | tam45 nt 1122184–1123706 in pCRA725; plasmid for allelic replacement of CgR_1015-1016 | XbaI/SalI | This study |
| pCRE539 | GLY3 nt 1509110–1510281 in pCRA725; plasmid for allelic replacement of CgR_1377-1378 | XbaI/SalI | This study |
| pCRE540 | tam45 nt 1509110–1510281 in pCRA725; plasmid for allelic replacement of CgR_1377-1378 | XbaI/SalI | This study |
| pCRE541 | GLY3 nt 1552995–1554627 in pCRA725; plasmid for allelic replacement of CgR_1414-1415 | SacI/SalI | This study |
| pCRE542 | tam45 nt 1552995–1554627 in pCRA725; plasmid for allelic replacement of CgR_1414-1415 | SacI/SalI | This study |
| pCRE543 | GLY3 nt 1614878–1617235 in pCRA725; plasmid for allelic replacement of CgR_1462-1463 | SacI/XbaI | This study |
| pCRE544 | tam45 nt 1614878–1617235 in pCRA725; plasmid for allelic replacement of CgR_1462-1463 | SacI/XbaI | This study |
| pCRE545 | GLY3 nt 3283826–3285778 in pCRA725; plasmid for allelic replacement of CgR_2959-2960 | XbaI/SalI | This study |
| pCRE546 | tam45 nt 3283826–3285778 in pCRA725; plasmid for allelic replacement of CgR_2959-2960 | XbaI/SalI | This study |
| pCRE552 | R nt 2556378–2572116 in pCRA725; plasmid for deletion of transgenes pfk and pyk | XbaI | This study |
| pCRE553 | R nt 791912–792959 fused to R nt 799755–800795 in pCRA725; plasmid for deletion of region 2 | SalI | This study |
| pCRE554 | R nt 2554655–2555773 fused to R nt 2560444–2561451 in pCRA725; plasmid for deletion of region 1 | SalI | This study |
| pCRE555 | Coding region of pyk cloned into pCRB214 | NdeI | This study |
| pCRE556 | Ptac-pyk-rrnBT1T2 cloned into pCRB12i | BamHI | This study |
| pCRE557 | Coding region of pfk cloned into pCRB214 | NdeI | This study |
| pCRE558 | Ptac-pfk-rrnBT1T2 cloned into pCRE556 | KpnI | This study |
a
Restriction site(s) used for cloning. NA, not applicable.
b
nt, nucleotides.