Table 2.
E. coli strains1, plasmids, and primers used in this work
| Strain/plasmid/primer | Description | Source or reference |
| Strains | ||
| BE3471 | PS2209 gltB (psiQ39)::lacZ (Mu d1-1734) | [28] |
| BE3479 | PS2209 gltB (psiQ32)::lacZ (Mu d1-1734) | [28] |
| BE3779 | PS2209 gltB (psiQ32)::lacZ (Mu d1-1734) lrp-35::Tn10 | [28] |
| BL21(DE3) | F- ompT hsdSB (rB-mB-) gal dcm (DE3) | Novagen |
| JWD3 | BE1 (W3110 lrp-201::Tn10) pJWD2 | [28] |
| LP1000 | PS2209 gltB::lacZ transcriptional fusion at λatt with -406 to +246 of the gltBDF operon | [14] |
| LP1050 | LP1000 ΔargR | This work |
| LP1060 | BL21DE3/pLP1060 | This work |
| LP1070 | BL21DE3/pLP1070 | This work |
| LP1080 | BL21DE3/pLP1080 | This work |
| LP1270 | PS2209 gltB::lacZ transcriptional fusion at λatt with -270 to +246 of the gltBDF operon | This work |
| LP2002 | W3110 rph+ | This work |
| LP2010 | LP2002 ΔlacZYA | This work |
| LP2020 | LP2010 gltB::lacZ transcriptional fusion at λatt with -406 to +246 of the gltBDF operon | This work |
| LP2023 | LP2020 ΔargR | This work |
| PM2003 | PS2209/pPM2005 | This work |
| PM2004 | PS2209/pPM2006 | This work |
| PS2209 | W3110 Δlac-169 | F. C. Neidhardt |
| W3110 | F- prototroph rph | F. C. Neidhardt |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 Δlac-pro [F' proAB lacIqZΔM15 Tn10] | Stratagene |
| Plasmids | ||
| pBH403 | pKK232-8 with promoterless lacZ gene inserted upstream of cat gene in a derivative of pBR322 | [61, 62] |
| pPM2005 | pBH403 with gltB promoter region cloned from BamHI to SalI | This work |
| pPM2006 | pPM2005 with mutations in the Crp-binding sequence | This work |
| PJWD2 | pTrc99A (Pharmacia) with the lrp coding sequence cloned into the NcoI site | [28] |
| pLP1060 | pET23b (Novagen) with the argR coding sequence cloned into the NdeI/XhoI sites | This work |
| pLP1070 | pET29a (Novagen) with the crp coding sequence inserted into the NdeI/XhoI sites | This work |
| pLP1080 | pET28A (Novagen) with the himA and himD coding sequences inserted in tandem into the NcoI//BamHI sites | This work |
| pGEMT-easy | Cloning vector | Promega |
| pKO3 | Suicide vector | [54] |
| Primers | ||
| arg1 | CGCCAGCAGCGCCGAGGACTGCGAC | |
| arg2 | CACTAATTATTGAGCTAATTAATACCGCGC | |
| arg3 | CATATGCGAAGCTCGGCTAAG | |
| arg4 | CTCGAGTTATTAAAGCTCCTGGTCGAACAGCTC | |
| crp5 | CCATATGGTGCTTGGCAAACCGC | |
| crp6 | CCTCGAGTTAACGAGTGCCGTAAACGAC | |
| gltP1 | CGGGATCCCATAATCACATAAATCACTTTTGCTTATC | |
| gltP2 | ACGCGTCGACAGCGGATTTCCAACTTATCG | |
| gltcrp12 | CAGTCAATTAATAAAGAATATAACGCTAAAGGCGGTTTCTGTACCAATAAGCTTGCC | |
| gltcrp2 | GGCAAGCTTATTGGTACAGAAACCGCCTTTAGCGTTATATTCTTTATTAATTGACTG | |
| himA1 | GGTACCGGCATCATTGAGGGATTGAACCTATGGCGCTTACA | |
| himA2 | CTCGAGCTCGTCTTTGGGCGAAGC | |
| himD1 | GGTACCTTAACCGTAAATATTGGCGCGATCGCG | |
| himD2 | TCATGACCAAGTCAGAATTGATAGAAGAC | |
| lac6 | GGATACGACGATACCGAAGACAGCTCATG | |
| lac7 | CGAGCCGGAAGCATAAAGTGTAAAGC | |
| lac8 | GCTTTACACTTTATGCTTCCGGCTCGGGCGTTCCTTGTCGGGTTATTCG | |
| rph1 | GCGTCTGATCGCCCGTGCTCTTCGCGC | |
| rph23 | CGTCGCTACAATGGATTCGATTCCCCCTCGGGC | |
| rph33 | GCCCGAGGGGGAATCGAATCCATTGTAGCGACG | |
| rph4 | ACGGCAGGTCCAGGTCGTGATGCTCCG | |
1All strains are Escherichia coli K-12.
2The underlined bases alter the Crp-binding site in the gltBDF promoter.
3The underlined italic base is inserted to replace the base deleted in the rph mutation.