Isolation of cDNA encoding the catalytic site of phosphatidylinositol-specific phospholipase C from Coffea arabica L.: Recombinant expression and peptide purification.
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.
Sánchez-Cach
Lucila A
LA
Unidad de Bioquímica y Biología Molecular de Plantas; Centro de Investigación Científica de Yucatán; Mérida, Yucatán México.
Ortiz-García
Matilde M
MM
Minero-García
Yereni
Y
Muñoz-Sánchez
J Armando
JA
Hernández-Sotomayor
Sm Teresa
ST
Suárez-Solís
Víctor M
VM
De Los Santos-Briones
César
C