TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| R. eutropha strains | ||
| CH34 | Wild type, containing pMOL28 and pMOL30, czc cnr chr mer cop pbr | 15 |
| AE126 | pMOL28, cnr chr mer | 15 |
| AE104 | Plasmid free, metal sensitive | 15 |
| AE963 | pMOL29, cnrY963::IS1087 chr mer, ZinB phenotype (due to derepression of cnr) | 1 |
| E. coli strains | ||
| DH10B | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 endA1 araD139 Δ(ara-leu)7697 galU galK λ− rpsL nupG | GIBCO BRL |
| CM1446 | DH10B containing pMOL877 | 30 |
| JM105 | F′ traD36 lacIqΔ(lacZ)M15 proA +B+/thi rpsL (Strr) endA sbcB15 sbcC hsdR4 (rK− mK+) Δ(lac-proAB) | 32 |
| BL21(DE3) | F−dcm ompT hsdS (rB− mB−) galDE3 | Stratagene |
| TOP10 F′ | F′ mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| DH5α | F− φ80d lacZΔM15 Δ(lacZyA-argF)U169 deoR recA1 endA1 phoA hsdR17 (rK− mK+) supE44 λ−thi-1 gyrAg6 relA1 | GIBCO BRL |
| pMOL877-based plasmids | ||
| pMOL877 | luxCDABE, Tetr | 30 |
| pMOL1550 | cnrYXH-luxCDABE, Tetr | This study |
| pMOL1551 | cnrYXHC′-luxCDABE, Tetr | This study |
| pMOL1583 | cnrYp-luxCDABE, Tetr | This study |
| pMOL1561 | cnrYXa H-luxCDABE, Tetr | This study |
| pMOL1586 | cnrYp-cnrYX-cnrHp-luxCDABE, Tetr | This study |
| pMOL1587 | cnrHp-luxCDABE, Tetr | This study |
| pMOL1588 | cnrHp-cnrH-luxCDABE, Tetr | This study |
| pMOL1591 | cnrX-cnrHp-luxCDABE, Tetr | This study |
| pMOL1592 | cnrYp-cnrY-luxCDABE, Tetr | This study |
| pMOL1593 | (cnrY-cnrYp) -cnrX-cnrHp-luxCDABEb, Tetr | This study |
| pMOL1596 | ivrHC-luxCDABE, Tetr | This study |
| Other plasmids | ||
| pTrc99A | Ampr | Pharmacia Biotech Ltd. |
| pTrcCnrX | Ampr, cnrX in pTrc99A | This study |
| pTrcCnrYa | Ampr, cnrYa in pTrc99A | This study |
| pTrcCnrYb | Ampr, cnrYb in pTrc99A | This study |
| pCnrXphoA1 | Ampr Kmr, pTrc-cnrX′ in pGV4218 | This study |
| pCnrXphoA2 | Ampr Kmr, cnrX′-pTrc in pGV4218 | This study |
| pGV4218 | Ampr Kmr | 5, 19 |
| pVDZ′2-cnr | cnr as EcoRI-PstI fragment in pVDZ′2 | 13 |
| pRSETA | Ampr | Invitrogen |
| pRSETCnrX | Ampr, cnrX in pRSETA | This study |
| pRSETCnrH | Ampr, cnrH in pRSETA | This study |
| pMOL29 | pMOL28 containing cnrY963::IS1087 | 1 |
a
Point mutations in PH region.
b
cnrY-cnrYp region cloned in the opposite orientation as cnrX-cnrHp-luxCDABE.