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Plant and Cell Physiology, 2003, Vol. 44, No. 2 139-145
© 2003 Oxford University Press

Characterization and Expression of a Novel Member (JBURE-II) of the Urease Gene Family from Jackbean [Canavalia ensiformis (L.) DC]

Melissa Pires-Alves1, Maria Fátima Grossi-de-Sá2, Grace B. S. Barcellos5, Célia R. Carlini1,3,6 and Marcelo G. Moraes4

1 Graduate Program in Cellular and Molecular Biology, Center of Biotechnology, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves no. 9500, Porto Alegre, RS, 91501-970, Brazil
2 Cenargen, Embrapa, SAIN, Parque Rural, Caixa Postal 02372, 70770-900, Brasília, DF, Brazil
3 Department of Biophysics, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves no. 9500, Porto Alegre, RS, 91501-970, Brazil
4 Department of Plant Pathology and Entomology, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves no. 7712, Porto Alegre, RS, 91540-000, Brazil

Canavalia ensiformis (jackbean) seeds contain the proteins urease and canatoxin, a variant form of the jackbean urease. Here we have cloned a cDNA encoding another isoform of urease, called JBURE-II. This cDNA was obtained by RT-PCR using as template total RNA extracted from C. ensiformis tissues. Nucleotide sequence analysis showed that JBURE-II clones share 86% similarity with known jackbean urease. The presence in C. ensiformis of a family of urease-related genes with at least three members was demonstrated by Southern blot analysis. In order to understand the pattern of expression of the JBURE-II gene, we collected tissue samples from different stages of flower and embryo development. The results of RT-PCR show that JBURE-II is expressed from flower buds throughout seed maturation. Semi-quantitative RT-PCR indicates that expression of urease and JBURE-II genes is induced in seedlings and in leaves treated with abscisic acid, a phytohormone involved in seed maturation and wound response. This work constitutes the first report on the presence of a family of urease genes in jackbean, and provides characterization of a cDNA encoding a new member of this gene family.

5 In memorium.

6 Corresponding author: E-mail, ccarlini{at}ufrgs.br; Fax, (55) (51) 3316-7003.


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