Cloning, mutagenesis, and nucleotide sequence of a siderophore biosynthetic gene (amoA) from Aeromonas hydrophila
| Autor: | Jean E. L. Arceneaux, S. Barghouthi, Shelley M Payne, B. R. Byers |
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| Rok vydání: | 1991 |
| Předmět: |
Molecular Sequence Data
Restriction Mapping Mutant Siderophores Biology Molecular cloning Iron Chelating Agents medicine.disease_cause Microbiology Enterobactin chemistry.chemical_compound Operon Escherichia coli Hydroxybenzoates medicine Genomic library Amino Acid Sequence Cloning Molecular Isomerases Intramolecular Transferases Molecular Biology Peptide sequence Gene Base Sequence Nucleic acid sequence chemistry Biochemistry Mutagenesis Aeromonas Oligopeptides Research Article |
| Zdroj: | Journal of Bacteriology. 173:5121-5128 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.173.16.5121-5128.1991 |
| Popis: | Many isolates of the Aeromonas species produce amonabactin, a phenolate siderophore containing 2,3-dihydroxybenzoic acid (2,3-DHB). An amonabactin biosynthetic gene (amoA) was identified (in a Sau3A1 gene library of Aeromonas hydrophila 495A2 chromosomal DNA) by its complementation of the requirement of Escherichia coli SAB11 for exogenous 2,3-DHB to support siderophore (enterobactin) synthesis. The gene amoA was subcloned as a SalI-HindIII 3.4-kb DNA fragment into pSUP202, and the complete nucleotide sequence of amoA was determined. A putative iron-regulatory sequence resembling the Fur repressor protein-binding site overlapped a possible promoter region. A translational reading frame, beginning with valine and encoding 396 amino acids, was open for 1,188 bp. The C-terminal portion of the deduced amino acid sequence showed 58% identity and 79% similarity with the E. coli EntC protein (isochorismate synthetase), the first enzyme in the E. coli 2,3-DHB biosynthetic pathway, suggesting that amoA probably encodes a step in 2,3-DHB biosynthesis and is the A. hydrophila equivalent of the E. coli entC gene. An isogenic amonabactin-negative mutant, A. hydrophila SB22, was isolated after marker exchange mutagenesis with Tn5-inactivated amoA (amoA::Tn5). The mutant excreted neither 2,3-DHB nor amonabactin, was more sensitive than the wild-type to growth inhibition by iron restriction, and used amonabactin to overcome iron starvation. |
| Databáze: | OpenAIRE |
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