TABLE 1.
Bacteria and plasmids involved in this study
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Bacteria | ||
| Corynebacterium glutamicum | ||
| ATCC 13032 | Wild type, plasmid free, Nxr | ATCC |
| RES167 | Restriction-deficient mutant of ATCC 13032, Δ(cglIM-cglIR-cglIIR) | 55 |
| Escherichia coli | ||
| DH5α | λ− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | Gibco BRL |
| Rosetta(DE3)pLysS | Camr, F− ompT hsdSB(rB− mB−) gal dcm (DE3), pLysSRARE vector (Camr) | Novagen |
| Trans1-T1 phage resistant | F− ϕ80(lacZ)ΔM15 ΔlacX74 hsdR(rK− mK+)ΔrecA1398 endA1 tonA | TransGen Biotech |
| Plasmids | ||
| pMD18-T | Ampr, lacZα, cloning vector for sequencing | TaKaRa |
| pET-28a(+) | Expression vector, Kanr, C/N-terminal His tag/thrombin/T7 tag, T7 lac promoter, T7 transcription start, f1 origin, lacI | Novagen |
| pZWHJ024 | Kanr, 681-bp PCR fragment of glxR gene ORF in C. glutamicum ATCC 13032 genome cloned into pET-28a(+) NdeI/EcoRI site, His tag banding C-terminal GlxR | This study |
| pZWHJ002 | Camr, pBL1 oriVCg, pK18, oriVEc, trrnB of E. coli, lacZ promoterless, derivative of pXMJ19 with lacZ digested with the enzyme combination NarI/HindIII, deletion of Ptac lacq, shuttle vector | 25 |
| pZWHJ006 | Camr, derivative of pZWHJ002 with 463-bp promoter of genDFM operon for detecting promoter activation | 25 |
| pZWHJ007 | Camr, derivative of pZWHJ002 with 457-bp promoter of genR operon for detecting promoter activation | 25 |
| pZWHJ008 | Camr, derivative of pZWHJ002 with 427-bp promoter of genKH operon for detecting promoter activation | 25 |
| pZWHJ025 | Camr, pZWHJ006 with site DFMx mutated | This study |
| pZWHJ026 | Camr, pZWHJ007 with site R-KHx01 mutated | This study |
| pZWHJ027 | Camr, pZWHJ007 with site R-KHx02 mutated | This study |
| pZWHJ028 | Camr, pZWHJ007 with sites R-KHx01 and R-KHx02 mutated | This study |
| pZWHJ029 | Camr, pZWHJ008 with site R-KHx01 mutated | This study |
| pZWHJ030 | Camr, pZWHJ008 with site R-KHx02 mutated | This study |
| pZWHJ031 | Camr, pZWHJ008 with sites R-KHx01 and R-KHx02 mutated | This study |
| pZWHJ033 | Camr, pBL1 oriVCg, pK18, oriVEc, trrnB of E. coli, carrying glxR gene, derivative of pXMJ19 with glxR from RES167 chromosome, digested with the enzyme combination SalI and EcoRI, shuttle vector | This study |
a
oriVCg, C. glutamicum oriV; oriVEc, E. coli oriV.