To focus the evaluation of protein profiles of the Acetic Acid Bacteria (AAB) symbionts by suitable protein extraction methods in order to perform two-dimensional gel electrophoresis (2DE). Good quality of protein extraction of all strains (AAB) were obtained using SDS-PAGE. Quantification of protein were performed by Bradford method to measure the amount of protein concentration present in the cell pellet and in parallel performed the SDS-PAGE (Urea sample) as control. Based on the protein quality and quantity of all strains, A. indonesiensis and Asaia SF2.1 were selected for 2D-PAGE setup. Three different trials of sample preparation methods like Urea Buffer extract, NaOH and TCA-Acetone were performed for 2D-PAGE setup. Compared to all methods, TCA-Acetone were selected as one of the best protein precipitation method for 2D-PAGE. Further, program setup were optimized and obtained the clear spots of protein profile of Asaia SF2.1. Finally, experiments will be done for comparative proteomic analysis of the selected bacterial strains in different growth conditions.
|Titolo:||Interactions between microbes and host of agricultural and environmental interest|
|Data di pubblicazione:||19-dic-2014|
|Parole Chiave:||Acetic Acid Bacteria; 2D-PAGE; TCA-Acetone|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore AGR/16 - Microbiologia Agraria
|Citazione:||Interactions between microbes and host of agricultural and environmental interest / S. Santiagu. ((Intervento presentato al 10. convegno Workshop : Doctorate in Chemistry, Biochemistry and Ecology of Plant Protection Products and Xenobiotics tenutosi a Milano nel 2014.|
|Appare nelle tipologie:||14 - Intervento a convegno non pubblicato|