TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | F′ endA1 hsdR17 (rk− mk+) supE44 thi-1 recA1 gyrA (Nalr) relA1 Δ(lacIZYA-argF)U169 deoR (φ80dlacΔ(lacZ)M15), host cells for all plasmid constructs | Michael S. DuBow |
| M15 | F′ lacIq Δ(lacZ)M15 supE44 lacY1 lacZ ara-14 galK2 xyl-5 mtl-1 leuB6 proA2 Δ(mcrC-mrr) recA+rpsL20 thi-1 λ− | Michael S. Dubow |
| BL21(DE3) | B F−ompT hsdS(rB− mB−) dcm+ Tetrgal 1 (DE3) endA Hte | Stratagene |
| CodonPlus RIL | [argU ileY leuW Camr] | |
| GM2589 | pGex 2T::CtrA in E. coli M15 | This study |
| GM2773 | pTrcHisA::CzcR in E. coli BL21(DE3) CodonPlus RIL | This study |
| K5746 | pPLhip.himA-5 in E. coli N5271 (IHF overexpressing strain) | 25 |
| C. crescentus strains | ||
| SC1107 | CB15N Δbla::Tn5, AmpS | 12 |
| LS1094 | ctrA401 (Ts) | 29 |
| GM2785 | pUJ142 in LS1094 | This study |
| GM2786 | pGM2456 in LS1094 | This study |
| GM2787 | pGM2767 in LS1094 | This study |
| Plasmids | ||
| pGM976 | pRKlac290 Cori BamHI to Ps::lacZ | 23 |
| pGM2656 | pACYC177 BglII PL+, Ampr | Lucille Shapiro |
| pGM2657 | pACYC177 BglII PL− Ampr | Lucille Shapiro |
| pGM2667 | pGM2656 EcoRI/SalI oriRp from pDW1070 | This study |
| pGM2668 | pGM2657 SalI/EcoRI oriRp from pDW1070 | This study |
| pGM2669 | pGM2656 EcoRI/SalI large fragment of oriRp from pDW1095 | This study |
| pGM2670 | pGM2657 SalI/EcoRI large fragment of oriRp from pDW1095 | This study |
| pGM2671 | pGM2656 BamHI/BamHI Cori from pGM1500 | This study |
| pGM2672 | pGM2657 BamHI/BamHI Cori from pGM1500 | This study |
| pAGMT* | pACYC184 derivative with RK2 oriT, Gmr Tcs | 23 |
| pGM2703 | SalI/BamHI oriRp from pDW1070 in pAGMT* | This study |
| pGM2704 | SalI/BamHI large fragment of oriRp from pDW1095 in pAGMT* | This study |
| pAGMT*E | pAGMT* BamHI fragment E from cosmid I | 23 |
| pUJ142 | Xylose-inducible promoter cloning vector, Cmr | Urs Jenal |
| pGM2767 | pUJ142::czcR | This study |
| pGM2456 | pUJ142::ctrA | This study |
| pBluescript(II) | SK(+) cloning vector, Ampr | Stratagene |
| pGM1500 | Cori BamHI/BamHI in pSK(+) | 23 |
| pGM1877 | Cori HindIII at position 214 to XhoI in pSK(+) | 33 |
| pMW1070 | PCR 492-bp oriRp blunt ligated into EcoRV site in pSK(+) | This study |
| pMW1089 | PCR 758-bp czcR blunt ligated into SmaI with engineered NdeI and BamHI sites of pSK(+) | This study |
| pMW1095 | PCR 1,753-bp large fragment containing oriRp blunt ligated into EcoRv in pSK(+) | This study |