PVAS3, a class-II ubiquitous asparagine synthetase from the common bean (Phaseolus vulgaris).
A gene encoding a putative asparagine synthetase (AS; EC 6.3.5.4) has been isolated from common bean (Phaseolus vulgaris). A 2.4 kb cDNA clone of this gene (PVAS3) encodes a protein of 570 amino acids with a predicted molecular mass of 64,678 Da, an isoelectric point of 6.45, and a net charge of -5.9 at pH 7.0. The PVAS3 protein sequence conserves all the amino acid residues that are essential for glutamine-dependent AS, and PVAS3 complemented an E. coli asparagine auxotroph, that demonstrates that it encodes a glutamine-dependent AS. PVAS3 displayed significant similarity to other AS. It showed the highest similarity to soybean SAS3 (92.9% identity), rice AS (73.7% identity), Arabidopsis ASN2 (73.2%) and sunflower HAS2 (72.9%). A phylogenetic analysis revealed that PVAS3 belongs to class-II asparagine synthetases. Expression analysis by real-time RT-PCR revealed that PVAS3 is expressed ubiquitously and is not repressed by light.
Parra-Peralbo
Esmeralda
E
Departamento de Botánica, Ecología y Fisiología Vegetal, Instituto Andaluz de Biotecnología, Universidad de Córdoba, Campus de Rabanales, Edif. C-4, 3a Planta, 14071 Córdoba, Spain.
Pineda
Manuel
M
Aguilar
Miguel
M
D000595:Amino Acid Sequence; D001217:Aspartate-Ammonia Ligase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D017343:Genes, Plant; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D027805:Phaseolus / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment; D014018:Tissue Distribution
D001217:Aspartate-Ammonia Ligase