TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Marker(s)a | Relevant genotype | Relevant characteristics and use | Reference or sourceb |
|---|---|---|---|---|
| Halobacterium strains | ||||
| MPK414 | Δura3 | Able to synthesize Cbl de novo | 50 | |
| JE6738 | Δura3 ΔcbiP | Unable to synthesize cobamides de novo | 51 | |
| JE7084 | Δura3 ΔcbiP ΔbtuCD | In-frame deletion of btuCD; unable to synthesize cobamides de novo | ||
| JE7108 | Δura3 ΔbtuCD | In-frame deletion of btuCD | ||
| JE7494 | Δura3 ΔcbiP ΔbtuCD ura3::cbiP+ | Complementation of de novo pathway | ||
| JE7495 | Δura3 ΔcbiP ΔbtuCD ura3::btuCD+ | Complementation of btuCD | ||
| JE7681 | Δura3 ΔbtuF | In-frame deletion of btuF | ||
| JE7682 | Δura3 ΔcbiP ΔbtuF | In-frame deletion of btuF; unable to synthesize cobamides de novo | ||
| JE7683 | Δura3 ΔcbiP ΔbtuF ΔbtuCD | In-frame deletions of btuF and btuCD; unable to synthesize cobamides de novo | ||
| JE7684 | Δura3 ΔcbiP ΔbtuCD ura3::btuC+ | Complementation of btuC | ||
| JE7685 | Δura3 ΔcbiP ΔbtuCD ura3::btuD+ | Complementation of btuD | ||
| JE7796 | Δura3 ΔcbiP ΔbtuF ura3::cbiP+ | Complementation of de novo pathway in btuF mutant | ||
| JE7797 | Δura3 ΔcbiP ΔbtuF ura3::btuF+ | Complementation of btuF | ||
| S. enterica strains | ||||
| JE873 | metE205 ara-9 Δcob272 (cobUST) | Cobamide bioassay | Laboratory collection | |
| JE2243 | metE205 ara-9 btuB101::MudJc | Cobamide bioassay | Laboratory collection | |
| Plasmids | ||||
| pMPK428 | 5-FOAs Mevr | ura3+ | Generation of in-frame deletions of targeted genes | 32 |
| pMPK424 | 5-FOAs Mevr | ura3+ | Recombination at ura3 locus | 31 |
| pCBIP7 | 5-FOAs Mevr | ΔcbiP | Generates cbiP deletion | 51 |
| pVNG1370-2 | 5-FOAs Mevr | ura3+ ΔbtuCD | Generates btuCD deletion | |
| pVNG1370-4 | 5-FOAs Mevr | ura3+btuCD+ | Recombination of btuCD into ura3 locus | |
| pHsBTUC5 | 5-FOAs Mevr | ura3+btuC+ | Recombination of btuC into ura3 locus | |
| pHsBTUD1 | 5-FOAs Mevr | ura3+btuD+ | Recombination of btuD into ura3 locus | |
| pHsBTUF5 | 5-FOAs Mevr | ura3+ ΔbtuF | Generates btuF deletion | |
| pHsBTUF6 | 5-FOAs Mevr | ura3+btuF+ | Recombination of btuF into ura3 locus |
a
Abbreviations: Mevr, resistance to mevinolin; 5-FOAs, sensitivity to 5-FOA.
b
Unless otherwise stated, strains and plasmids were constructed during the course of this study.
c
Abbreviation of Mu dI1734 (10).