title: Putative virulence factors of Corynebacterium pseudotuberculosis FRC41: vaccine potential and protein expression.
abstract: BACKGROUND
  Corynebacterium pseudotuberculosis, a facultative intracellular bacterial pathogen, is the etiological agent of caseous lymphadenitis (CLA), an infectious disease that affects sheep and goats and it is responsible for significant economic losses. The disease is characterized mainly by bacteria-induced caseous necrosis in lymphatic glands. New vaccines are needed for reliable control and management of CLA. Thus, the putative virulence factors SpaC, SodC, NanH, and PknG from C. pseudotuberculosis FRC41 may represent new target proteins for vaccine development and pathogenicity studies.
  
  
  RESULTS
  SpaC, PknG and NanH presented better vaccine potential than SodC after in silico analyses. A total of 136 B and T cell epitopes were predicted from the four putative virulence factors. A cluster analysis was performed to evaluate the redundancy degree among the sequences of the predicted epitopes; 57 clusters were formed, most of them (34) were single clusters. Two clusters from PknG and one from SpaC grouped epitopes for B and T-cell (MHC I and II). These epitopes can thus potentially stimulate a complete immune response (humoral and cellular) against C. pseudotuberculosis. Several other clusters, including two from NanH, grouped B-cell epitopes with either MHC I or II epitopes. The four target proteins were expressed in Escherichia coli. A purification protocol was developed for PknG expression.
  
  
  CONCLUSIONS
  In silico analyses show that the putative virulence factors SpaC, PknG and NanH present good potential for CLA vaccine development. Target proteins were successfully expressed in E. coli. A protocol for PknG purification is described.
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, Pampulha, Belo Horizonte, 31270-901, Brazil.
authors.forename: Karina T O
authors.initials: KT
authors.lastname: Santana-Jorge
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, Pampulha, Belo Horizonte, 31270-901, Brazil.|Uniclon Biotecnologia, Belo Horizonte, MG, Brazil.
authors.forename: Túlio M
authors.initials: TM
authors.lastname: Santos
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, Pampulha, Belo Horizonte, 31270-901, Brazil.
authors.forename: Natayme R
authors.initials: NR
authors.lastname: Tartaglia
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, Pampulha, Belo Horizonte, 31270-901, Brazil.
authors.forename: Edgar L
authors.initials: EL
authors.lastname: Aguiar
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, Pampulha, Belo Horizonte, 31270-901, Brazil.
authors.forename: Renata F S
authors.initials: RF
authors.lastname: Souza
authors.affiliation: Multiuser Center for Biomolecular Innovation, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista "Júlio de Mesquita Filho", São José Do Rio Preto, SP, Brazil.
authors.forename: Ricardo B
authors.initials: RB
authors.lastname: Mariutti
authors.affiliation: Multiuser Center for Biomolecular Innovation, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista "Júlio de Mesquita Filho", São José Do Rio Preto, SP, Brazil.
authors.forename: Raphael J
authors.initials: RJ
authors.lastname: Eberle
authors.affiliation: Multiuser Center for Biomolecular Innovation, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista "Júlio de Mesquita Filho", São José Do Rio Preto, SP, Brazil.
authors.forename: Raghuvir K
authors.initials: RK
authors.lastname: Arni
authors.affiliation: Laboratório de Imunologia e Biologia Molecular, Instituto de Ciências da Saúde, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Ricardo W
authors.initials: RW
authors.lastname: Portela
authors.affiliation: Laboratório de Imunologia e Biologia Molecular, Instituto de Ciências da Saúde, Universidade Federal da Bahia, Salvador, BA, Brazil.
authors.forename: Roberto
authors.initials: R
authors.lastname: Meyer
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, Pampulha, Belo Horizonte, 31270-901, Brazil. vasco@icb.ufmg.br.
authors.forename: Vasco
authors.initials: V
authors.lastname: Azevedo
mesh_terms: D000595:Amino Acid Sequence; D001428:Bacterial Vaccines / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D016000:Cluster Analysis; D016925:Corynebacterium pseudotuberculosis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism / Q000472:pathogenicity*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D037521:Virulence Factors / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
chemical_list: D001428:Bacterial Vaccines; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D011994:Recombinant Proteins; D037521:Virulence Factors
keywords: Corynebacterium pseudotuberculosis; Epitope prediction; Pathogenicity and virulence; Protein expression; Protein purification; Vaccine potential