Title:	Identification of new Corynebacterium pseudotuberculosis antigens by immunoscreening of gene expression library.
Authors:	GALVÃO, C. E. FRAGOSO, S. P. OLIVEIRA, C. E. de FORNER, O. PEREIRA, R. R. B. SOARES, C. O. ROSINHA, G. M. S.
Affiliation:	CLEBER EDUARDO GALVÃO, Federal University of Mato Grosso do Sul (UFMS), Campo Grande, MS.; STENIO PERDIGÃO FRAGOSO, Carlos Chagas Institute, Oswaldo Cruz Foundation (ICC-FIOCRUZ), Curitiba, PR.; CARINA ELISEI DE OLIVEIRA, Dom Bosco Catholic University (UCDB), Campo Grande, MS.; ODINÉIA FORNER, Federal University of Paraná (UFPR), Curitiba, PR.; RENATA RIBEIRO BASTOS PEREIRA, Federal University of Mato Grosso do Sul (UFMS), Campo Grande, MS.; CLEBER OLIVEIRA SOARES, DE/TT; GRACIA MARIA SOARES ROSINHA, CNPGC.
Date Issued:	2017
Citation:	BMC Microbiology, v. 17, n. 202, p. 1-8, sept., 2017.
Description:	Background: Caseous lymphadenitis (CLA) is a disease that affects sheep, goats and occasionally humans. The etiologic agent is the Corynebacterium pseudotuberculosis bacillus. The objective of this study was to build a gene expression library from C. pseudotuberculosis and use immunoscreening to identify genes that encode potential antigenic proteins for the development of DNA and subunit vaccines against CLA. Results: A wild strain of C. pseudotuberculosis was used for extraction and partial digestion of genomic DNA. Sequences between 1000 and 5000 base pairs (bp) were excised from the gel, purified, and the digested DNA fragments were joined to bacteriophage vector ZAP Express, packaged into phage and transfected into Escherichia coli . For immunoscreening a positive sheep sera pool and a negative sera pool for CLA were used. Four clones were identified that strongly reacted to sera. The clones were confirmed by polymerase chain reaction (PCR) followed by sequencing for genomic comparison of C. pseudotuberculosis in GenBank. The genes identified were dak2, fagA, fagB, NlpC/P60 protein family and LPxTG putative protein family. Conclusion: Proteins of this type can be antigenic which could aid in the development of subunit or DNA vaccines against CLA as well as in the development of serological tests for diagnosis. Immunoscreening of the gene expression library was shown to be a sensitive and efficient technique to identify probable immunodominant genes.
Thesagro:	Corynebacterium Pseudotuberculosis
NAL Thesaurus:	Caseous lymphadenitis Sheep Gene expression
Keywords:	DNA vaccines
DOI:	10.1186/s12866-017-1110-7
Type of Material:	Artigo de periódico
Access:	openAccess
Appears in Collections:	Artigo em periódico indexado (CNPGC)