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Isolation of a complementary DNA encoding the bean PR4 chitinase: an acidic enzyme with an amino-terminus cysteine-rich domain
Authors:Marcia Margis-Pinheiro  Marie Hélène Metz-Boutigue  Abalo Awade  Marc de Tapia  Monique le Ret  Géraud Burkard
Affiliation:(1) Institut de Biologie Moléculaire des Plantes, 12, rue du général Zimmer, 67084 Strasbourg Cedex, France;(2) Institut de Biologie Moléculaire et Cellulaire, 15, rue Descartes, 67084 Strasbourg Cedex, France
Abstract:The amino acid sequences of peptides generated by trypsin and chymotrypsin digestions of the acidic PR4 chitinase from bean were determined. Oligonucleotide primers derived from this sequence were used to synthesize a PR4 chitinase-specific probe by PCR-amplification. This probe allowed the isolation of cDNA clones encoding PR4 chitinase that have been sequenced. This acidic and extracellular chitinase shows some homology to the basic isoform from the same plant, and differs from other known acidic chitinases by the presence of an amino-terminal cysteine-rich domain. Southern blot analysis of bean genomic DNA revealed that PR4 chitinase is encoded by a single gene.
Keywords:bean chitinase cDNA  cysteine-rich domain  plant defence  chemical stress
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