title: Isolation of cDNA encoding the catalytic site of phosphatidylinositol-specific phospholipase C from Coffea arabica L.: Recombinant expression and peptide purification.
abstract: A cDNA encoding the catalytic site of a phosphatidylinositol-specific phospholipase C (PI-PLC) was isolated from Coffea arabica suspension cells. The cDNA (designated CaPLC) encodes a polypeptide of 308 amino acids, containing the catalytic X and Y domains, and has 99% identity to the soybean gene. Recombinant CaPLC protein was expressed in Escherichia coli, purified, and used to produce a polyclonal antibody. The peptide has a molecular mass of 27 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot analyses. Immunoblots revealed the presence of PLC-like proteins in the tissues of different plant species.
authors.affiliation: Unidad de Bioquímica y Biología Molecular de Plantas; Centro de Investigación Científica de Yucatán; Mérida, Yucatán México.
authors.forename: Lucila A
authors.initials: LA
authors.lastname: Sánchez-Cach
authors.forename: Matilde M
authors.initials: MM
authors.lastname: Ortiz-García
authors.forename: Yereni
authors.initials: Y
authors.lastname: Minero-García
authors.forename: J Armando
authors.initials: JA
authors.lastname: Muñoz-Sánchez
authors.forename: Sm Teresa
authors.initials: ST
authors.lastname: Hernández-Sotomayor
authors.forename: Víctor M
authors.initials: VM
authors.lastname: Suárez-Solís
authors.forename: César
authors.initials: C
authors.lastname: De Los Santos-Briones