CBN-V Video Archives - S4-08
A Proteomic Approach to Isolate New Genes from Cassava Storage Roots

Rocha T.L. and Carvalho L.J.C.B.

EMBRAPA-Genetic Resources and Biotechnology, CP 02372, CEP 70770-900 Sain Parque Rural, Brazilia, Brazil carvalho@cenargen.embrapa.br

        Cassava (Manihot esculenta Crantz) plants are being used to apply a PROTEOM approach to isolate new genes from its storage root that are related to starch, carotenoid biosynthesis, and SR specific genes that are developmentally regulated. Three classes of SR clones of cassava were sorted in our genetic bank to represent diversity in the above traits as well as three SR tissue system in the compartmentalization model. Phosphate saline buffer protein extracts were fractionated in two affinity chromatographies (Con-A and Hydrophobic resins). Fractionated proteins are analyzed by SDS-PAGE, 2-DE gel, computer assisted analysis (GELLAB-IIa software), MALDI-TOF characterization of interest proteins, and finally protein sequencing. Soluble protein content varies among the classes of cassava clones, being high (2.8mg/gDWt) in the pigmented SR. SDS-PAGE protein profiles revealed a more complex protein population in the pigmented clones in relation to the non pigmented ones. Protein spots in 2-DE gels varying in pI and MW are being used to generate a 2-DE gel map for the tissue compartmentalization model and the genotypes used. Further characterization are being done with reduced number of interest protein for sequencing.

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