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Updated: Feb 15, 2026

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
Published on: January 6, 2015
Identification of membrane-associated proteins with pathogenic potential expressed by Corynebacterium
José Tadeu Raynal1, Bruno Lopes Bastos2, Priscilla Carolinne Bagano Vilas-Boas1
1Laboratório de Imunologia e Biologia Molecular (LABIMUNO), Departamento de Biointeração, Instituto de Ciências da Saúde (ICS), Universidade Federal da Bahia (UFBA), Av. Reitor Miguel Calmon, S/N, Vale do Canela, Salvador, BA, CEP 40140-100, Brazil.
Objective:
Previous works defining antigens that might be used as vaccine targets against Corynebacterium pseudotuberculosis, which is the causative agent of sheep and goat caseous lymphadenitis, have focused on secreted proteins produced in a chemically defined culture media. Considering that such antigens might not reflect the repertoire of proteins expressed during infection conditions, this experiment aimed to investigate the membrane-associated proteins with pathogenic potential expressed by C. pseudotuberculosis grown directly in animal serum.
Results:
Its membrane-associated proteins have been extracted using an organic solvent enrichment methodology, followed by LC-MS/MS and bioinformatics analysis for protein identification and classification. The results revealed 22 membrane-associated proteins characterized as potentially pathogenic. An interaction network analysis indicated that the four potentially pathogenic proteins ciuA, fagA, OppA4 and OppCD were biologically connected within two distinct network pathways, which were both associated with the ABC Transporters KEGG pathway. These results suggest that C. pseudotuberculosis pathogenesis might be associated with the transport and uptake of nutrients; other seven identified potentially pathogenic membrane proteins also suggest that pathogenesis might involve events of bacterial resistance and adhesion. The proteins herein reported potentially reflect part of the protein repertoire expressed during real infection conditions and might be tested as vaccine antigens.
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