PMID:2448295
| Citation |
Castaño, I, Bastarrachea, F and Covarrubias, AA (1988) gltBDF operon of Escherichia coli. J. Bacteriol. 170:821-7 |
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| Abstract |
A 2.0-kilobase DNA fragment carrying antibiotic resistance markers was inserted into the gltB gene of Escherichia coli previously cloned in a multicopy plasmid. Replacement of the chromosomal gltB+ gene by the gltB225::omega mutation led to cells unable to synthesize glutamate synthase, utilize growth rate-limiting nitrogen sources, or derepress their glutamine synthetase. The existence of a gltBDF operon encoding the large (gltB) and small (gltD) subunits of glutamate synthase and a regulatory peptide (gltF) at 69 min of the E. coli linkage map was deduced from complementation analysis. A plasmid carrying the entire gltB+D+F+ operon complemented cells for all three of the mutant phenotypes associated with the polar gltB225::omega mutation in the chromosome. By contrast, plasmids carrying gltB+ only complemented cells for glutamate synthase activity. A major tricistronic mRNA molecule was detected from Northern (RNA blot) DNA-RNA hybridization experiments with DNA probes containing single genes of the operon. A 30,200-dalton polypeptide was identified as the gltF product, the lack of which was responsible for the inability of cells to use nitrogen-limiting sources associated with gltB225::omega. |
| Links | |
| Keywords |
DNA, Bacterial/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Genes; Genes, Bacterial; Genetic Complementation Test; Glutamate Synthase/genetics; Glutamate Synthase/metabolism; Glutamate-Ammonia Ligase/genetics; Glutamate-Ammonia Ligase/metabolism; Mutation; Nucleic Acid Hybridization; Operon; Phenotype; Plasmids; RNA, Bacterial/genetics; Transaminases/genetics; Transcription Factors/genetics; Transcription, Genetic |
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