Characterization and expression of a novel member (JBURE-II) of the urease gene family from jackbean [Canavalia ensiformis (L.) DC |
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Authors: | Pires-Alves Melissa Grossi-de-Sá Maria Fátima Barcellos Grace B S Carlini Célia R Moraes Marcelo G |
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Institution: | Graduate Program in Cellular and Molecular Biology, Center of Biotechnology, Universidade Federal do Rio Grande do Sul, Av. Bento Gon?alves no. 9500, Porto Alegre, RS, 91501-970, Brazil. |
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Abstract: | 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. |
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