Molecular cloning and sequence analysis of an Azospirilium brasilense indole-3-pyruvate decarboxylase gene
| Autor: | Jos Vanderleyden, Antonia Costacurta, Veerle Keijers |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Carboxy-Lyases
Sequence analysis Recombinant Fusion Proteins Molecular Sequence Data Restriction Mapping Mutant Azospirillum brasilense Molecular cloning Biology chemistry.chemical_compound Bacterial Proteins Enterobacter cloacae Escherichia coli Genetics Amino Acid Sequence Cloning Molecular Molecular Biology Chromatography High Pressure Liquid Base Sequence Indoleacetic Acids Sequence Homology Amino Acid Nucleic acid sequence Indolepyruvate decarboxylase food and beverages Drug Resistance Microbial Sequence Analysis DNA biology.organism_classification Molecular biology Mutagenesis Insertional chemistry Biochemistry Genes Bacterial DNA Transposable Elements bacteria Indole-3-acetic acid Pyruvate decarboxylase Sinorhizobium meliloti |
| Zdroj: | Molecular and General Genetics MGG. 243:463-472 |
| ISSN: | 1432-1874 0026-8925 |
| DOI: | 10.1007/bf00280477 |
| Popis: | Azospirillum brasilense isolated from the rhizosphere of different plants has the ability to excrete indole-3-acetic acid (IAA) into the culture media. Cosmid p0.2, isolated from an A. brasilense Sp245 genome library in pLAFR1, complements the Tn5-induced mutant SpM7918 of A. brasilense Sp6 which excretes reduced amounts of IAA. Restriction mapping and gene expression studies identified a BglII-EcoRI 4.3 kb fragment of p0.2 sufficient for the restoration of high levels of IAA production in mutant SpM7918. Tn5 mutagenesis localized the gene responsible on a 1.8 kb SmaI fragment. Nucleotide sequence analysis revealed that this fragment contains one complete open reading frame. The predicted protein sequence shows extensive homology with the indole-3-pyruvate decarboxylase of Enterobacter cloacae and the pyruvate decarboxylases of Saccharomyces cerevisiae and Zymomonas mobilis. The A. brasilense mutant Sp245a, constructed by homogenotization of a Tn5 insertion derivative of the 1.8 kb SmaI fragment, also displayed reduced IAA production. Introduction of the cloned wild-type gene into Rhizobium meliloti 1021 resulted in increased IAA production. Cell-free extracts prepared from R. meliloti and A. brasilense transconjugants harboring this gene could convert indole-3-pyruvic acid to indole-3-acetaldehyde and tryptophol. These results clearly demonstrate that IAA production in A. brasilense is mediated by indole-3-pyruvate decarboxylase. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink