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PVAS3, a class-II ubiquitous asparagine synthetase from the common bean (Phaseolus vulgaris)

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Abstract

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.

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Acknowledgments

This work was funded by Dirección General de Enseñanza Superior (Grant BIO2006-09366) and by Plan Andaluz de Investigación (CV-0115). E.P.-P. thanks Félix Parra, Santi Peralbo for their teaching and continuous support, Minerva P–P for her invaluable technician help, and Joaquim Culí for his support and suggestions during the last stages of his work.

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  1. 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

    Esmeralda Parra-Peralbo, Manuel Pineda & Miguel Aguilar

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  1. Esmeralda Parra-Peralbo
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  2. Manuel Pineda
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  3. Miguel Aguilar
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Correspondence to Miguel Aguilar.

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Parra-Peralbo, E., Pineda, M. & Aguilar, M. PVAS3, a class-II ubiquitous asparagine synthetase from the common bean (Phaseolus vulgaris). Mol Biol Rep 36, 2249–2258 (2009). https://doi.org/10.1007/s11033-008-9441-y

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