INGEBI   02650
INSTITUTO DE INVESTIGACIONES EN INGENIERIA GENETICA Y BIOLOGIA MOLECULAR "DR. HECTOR N TORRES"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
GENOMIC AND IMMUNOLOGIC CHARACTERIZATION OF StCDPK1 AND STCDPK2
Autor/es:
VERÓNICA GIAMMARIA; PABLO GARGANTINI; SERGIO FEINGOLD; RITA M. ULLOA
Lugar:
Rosario, Santa Fé
Reunión:
Congreso; XLII reunión de SAIB; 2006
Institución organizadora:
SAIB
Resumen:
During tuberization, dormancy and sprouting of potato tubers, calcium-dependent protein kinase isoforms, StCDPK1 and StCDPK2, are differentially expressed. The genomic characterization of both isoforms was undertaken.  The complete genomic sequence of StCDPK1 was obtained and SSCP analysis was used to map the gene. Comparison of StCDPK1 with CDPK genes from Arabidopsis and rice genomes showed that it shares the 8-exons/7-introns structure of the CDPKs from group IIa. StCDPK1 maps in the distal end of chromosome 12, close to the TC69927 marker. A PCR approach was designed to amplify StCDPK2 gene assuming that both kinases share a similar gene structure. We were able to amplify the genomic sequence corresponding to the regulatory domain of StCDPK2. The results obtained show that intron´s length and positon are identical in both kinases. Experiments are being performed to obtain the complete genomic sequence and to map StCDPK2 gene in the potato genome. Western analysis and immunolocalization of StCDPK1 and StCDPK2 were performed. StCDPK1 was detected in the particulate fraction of stolons and histochemical analysis show that it is associated to the outer epidermal layer of cells while StCDPK2 was detected in soluble extracts. A recombinant StCDPK2-6xHis fusion protein was purified and enzymatic assays are being performed to characterize StCDPK2 kinetic parameters.