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Characterization of Mycoplasma agalactiae membrane proteome

Cacciotto, Carla (2011) Characterization of Mycoplasma agalactiae membrane proteome. Doctoral Thesis.

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Mycoplasma agalactiae is the etiological agent of contagious agalactia, a serious disease of small ruminants characterized by mastitis, polyarthritis, keratoconjunctivitis, and abortion. Mycoplasmas totally lack the cell wall, so membrane proteins are directly exposed to environment. To date, a limited number of M. agalactiae antigens have been described. The combination of 2-D PAGE and mass spectrometry is a well-established method for proteome studies; however, it is reported that standard 2-D PAGE lacks in resolution of hydrophobic and basic proteins, which are abundant in mycoplasmas membrane. Indeed, membrane proteins are poorly represented in total extracts maps. In this study, the membrane proteome of M. agalactiae PG2T was characterized. The Triton X-114 fractionation allowed the enrichment for M. agalactiae PG2T membrane proteins. Liposoluble proteins were subjected to 2-D PAGE-MS, leading to the identification of 40 unique proteins. The differential expression of liposoluble proteins, among PG2T strain and two field isolates, was revealed by 2D DIGE. The use of GeLC-MS/MS allowed to increase the coverage of the liposoluble proteome. A total of 194 unique proteins were identified, (26% of M. agalactiae PG2T genes) and subjected to gene ontology analysis for localization and function. Interestingly, the 11.5% of identified proteins derived from putative horizontal gene transfer events. This work paves the way for the development of diagnostic and prophylaxis tools.

Item Type:Doctoral Thesis
ID Code:5051
Contributors:Alberti, Alberto
Publisher:Universita' degli studi di Sassari
Uncontrolled Keywords:Mycoplasma, contagious agalactia, membrane, fractionation
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/05 Malattie infettive degli animali domestici
Divisions:001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
Cicli, scuole e corsi:Ciclo 23 > Scuole > Scienze biomolecolari e biotecnologiche > Microbiologia molecolare e clinica
Deposited On:01 Apr 2011 12:32

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