TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant characteristics or primer sequencea | Source or reference |
|---|---|---|
| E. coli strains | ||
| RP437 | Wild type reference strain for chemotaxis; F−thr-1 leuB6 his-4 metF159 thi-1 ara-14 lacY1 mtl-1 xyl-5 rpsL136 tonA31 tsx-78 eda-50 | 37 |
| RP1131 | RP437 trg-2::Tn10 | J. S. Parkinson |
| RP2361 | RP437 Δtar-386-2 | 32, 39 |
| RP3525 | RP437 Δtap-365-4 Δ(lac)U169 thr+leu+ | 32, 39 |
| RP5700 | RP437 Δtsr-7028 | J. S. Parkinson |
| UU1250 | RP437 Δaer-1 ygjG::Gm Δtsr-7028 Δ(tar-tap)5201 zbd::Tn5 Δtrg-100 thr+met+ | 9 |
| UU1615 | RP437 Δaer-1 ygjG::Gm Δ(tar-tap)5201 zbd::Tn5 Δtrg-100 thr+met+ | 4 |
| UU1624 | RP437 Δaer-1 ygjG::Gm Δtsr-7028 Δtap-3654 zbd::Tn5 Δtrg-100 thr+met+ | 4 |
| UU1625 | RP437 Δaer-1 ygjG::Gm Δtsr-7028 zbd::Tn5 Δtrg-100 | J. S. Parkinson |
| XL1 | RP437 ΔdppA::Km | This study |
| Plasmids | ||
| pRR48 | lacIq/ptac cloning vector, Apr | 42 |
| pXL2 | PCR fragment of tsr cloned into pRR48, Apr | This study |
| pXL3 | Chimeric fragment tsrap cloned into pRR48, Apr | This study |
| pXL4 | Chimeric fragment tapsr cloned into pRR48, Apr | This study |
| pXL5 | PCR fragment of tap cloned into pRR48, Apr | This study |
| pXL8 | Chimeric fragment tsrap plus the last 5 codons of tsr in pRR48, Apr | This study |
| pXL9 | Chimeric fragment tsrap plus the last 19 codons of tsr in pRR48, Apr | This study |
| pKD13 | Template plasmid, derivative of pANTSγ that contain an FRT-flanked kanamycin resistance gene | 12 |
| pKD46 | Red helper plasmid, derivative of pINT-ts which contain araC-ParaB and γ β exo | 12 |
| Primers | ||
| tsrNF | 5′-GCCGCCGCATTAATGTTAAAACGTATCAAAATTGTGACCAGC-3′ | This study |
| tsrNR | 5′-CCACGCAAAGCCTGCTGCATATGGCGCAAACTCTCTGCC-3′ | This study |
| tsrCF | 5′-GCCATTTTTGCCAGTCTGAAGCATATGCAGGGAGAGCTGATGC-3′ | This study |
| tsrCR | 5′-GCCGCGCCAAGCTTTAAAATGTTTCCCAGTTCTCCTCGC-3′ | This study |
| tapNF | 5′-CGCCGCATTAATGTTTAATCGTATTCGAATTTCGACCACGC-3′ | This study |
| tapNR | 5′-GCATCAGCTCTCCCTGCATATGCTTCAGACTGGCAAAAATGGC-3′ | This study |
| tapCF | 5′-GCAGAGAGTTTGCGCCATATGCAGCAGGCTTTGCGTGGGACGG-3′ | This study |
| tapCR | 5′-GCCGCGCCAAGCTTCAGGATACCACTGGCGCAATTTGTAACTGC-3′ | This study |
| tapCR5aa | 5′-GCCGCGCCAAGCTTCAGGATACCACTGGCGCAATTTGTAACTGC-3′ | This study |
| tapCRp | 5′-GGATACCACTGGCGCAATTTG-3′ | This study |
| 19aaFp | 5′-CCAGCTGCGCCGCGTAAAATGG-3′ | This study |
| 19aaR | 5′-GCCGCGCCAAGCTTTAAAATGTTTCCCAGTTC-3′ | This study |
| dppAH1P4 | 5′-AACAACAAACATCACAATTGGAGCAGAATAATGCGTATTCCGGGGATCCGTCGACC-3′ | This study |
| dppAH2P1 | 5′-GCCATCAGTCTTGTATGGCTTTTAATTATTCGATAGGTGTAGGCTGGAGCTGCTTC-3′ | This study |
a
Underlined bases indicate the positions of restriction sites.