Maserti Bianca Elena, Della Croce C.M., Luro François, Morillon Raphaël, Cini M., Caltavuturo L.. 2007. A general method for the extraction of citrus leaf proteins and separation by 2D electrophoresis : A follow up. Journal of Chromatography. B, 849 (1-2) : 351-356.
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Résumé : With the aim of studying differentially expressed proteins as a function of abiotic and biotic stress in citrus plants, we optimized a protocol for the extraction of total leaf proteins and their 2-DE separation using commercially available immobilized pH gradient strips (IPGs) in the first dimension. Critical factors for good reproducibility of citrus leaf protein separation were identified: trichloroacetic acid (TCA)/acetone precipitation after extraction in lysis buffer, sample fractionation on narrow range overlapping IPGs and sample-cup loading at the anodic or cathodic end of the strip. The use of thiourea and a strong detergent (C7BzO) in the solubilization/rehydration buffer, coupled with the increase to 10% of SDS in the equilibration buffer before the second dimension seemed to affect positively the resolution of basic proteins. Using our protocol we resolved about 30 basic proteins on 6.3-8.3 pH range strips. Further, our protocol was successfully applied reproducibly on the analysis of control and salt exposed leaf samples of Citrus reshni Hort. Ex Tan. © 2006 Elsevier B.V. All rights reserved.
Mots-clés Agrovoc : Citrus, protéine, extraction, séparation, électrophorèse, pH
Classification Agris : Q04 - Composition des produits alimentaires
Champ stratégique Cirad : Axe 3 (2005-2013) - Alimentation accessible et de qualité
Auteurs et affiliations
- Maserti Bianca Elena, CNR (ITA)
- Della Croce C.M., CNR (ITA)
- Luro François, INRA (FRA)
- Morillon Raphaël, CIRAD-FLHOR-UPR Multiplication végétative (FRA)
- Cini M., CNR (ITA)
- Caltavuturo L., CNR (ITA)
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Source : Cirad - Agritrop (https://agritrop.cirad.fr/539069/)
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