Article Figures & Data
Figures
- FIG. 1.
Structure of AdoCbl (coenzyme B12).
- FIG. 2.
Nutritional studies of Halobacterium sp. strain NRC-1 strains. Growth of Halobacterium sp. strain NRC-1 strains in the defined medium at 37°C is reported. (A and B) Growth in a liquid defined medium reported as CFU as a function of time. Strains are indicated by their genotype. Corrinoids added to the medium are indicated next to the genotype. (C, D, and E) Growth of cells seeded onto minimal agar plates and incubated for 8 days. Strains used were MPK414, cobY+; JE6736, ΔcobY; and JE6737 ΔcobY ura3::cobY+. Abbreviations: Cby, cobyric acid; Cbl, cobalamin. In all cases, corrinoids were added to 15 nM.
- FIG. 3.
Abbreviated view of the late steps of AdoCbl biosynthesis in bacteria (A) and archaea (B). Intermediates are boxed. Abbreviations: AP-Pi, aminopropanol phosphate; AdoCby, adenosylcobyric acid. (A) CbiB, putative AdoCbi-P synthase; CobU, AdoCbi:NTP kinase, GTP/AdoCbi-P guanylyltransferase; CobS, cobalamin (5′-P) synthase. (B) CobY, NTP:AdoCbi nucleotidyltransferase.
Tables
- TABLE 1.
Halobacterium sp. strain NRC-1 strains and plasmids used in this study
Strain Marker(s)a Relevant genotype Description Reference MPK414 Δura3 cobY+ Strain with de novo cobamide biosynthetic capability This study JE6736 Δura3 ΔcobY Strain with in-frame deletion of cobY This study JE6737 Δura3 ΔcobY ura3::cobY+ Strain used to test for complementation studies This study pMPK410 Mevr Δura3 Plasmid transformed into Halobacterium sp. strain NRC-1 to delete ura3 20 pMPK428 5-FOAs Mevr ura3 + Plasmid used to generate in-frame deletions of targeted genes 21 pCOBY30 5-FOAs Mevr ura3 + ΔcobY Plasmid transformed into MPK414 to delete cobY This study pMPK424 5-FOAs Mevr ura3 + Plasmid contains flanking sequence to ura3 to allow recombination at the chromosomal ura3 locus 22 pCOBY33 5-FOAs Mevr ura3 + cobY + Plasmid used to recombine cobY into ura3 locus This study ↵ a Abbreviations: Mevr, resistance to mevinolin; 5-FOAs, sensitivity to 5-fluoroorotic acid.
