Characterization and expression of a novel member (JBURE-II) of the urease gene family from jackbean [Canavalia ensiformis (L.) DC].
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.
Pires-Alves
Melissa
M
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.
Grossi-de-Sá
Maria Fátima
MF
Barcellos
Grace B S
GB
Carlini
Célia R
CR
Moraes
Marcelo G
MG
D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D001483:Base Sequence; D032449:Canavalia / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D006824:Hybridization, Genetic; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014510:Urease / Q000235:genetics* / Q000378:metabolism
D010940:Plant Proteins; D012333:RNA, Messenger; D000040:Abscisic Acid; D014510:Urease