期刊
BMC MICROBIOLOGY
卷 17, 期 -, 页码 -出版社
BMC
DOI: 10.1186/s12866-017-1110-7
关键词
Corynebacterium pseudotuberculosis; Caseous lymphadenitis; Sheep; Gene expression; Library
类别
资金
- Empresa Brasileira de Pesquisa Agropecuaria (Embrapa)
- Embrapa Goat and Sheep
- Northeast of Brazil Bank
- Higher Education Personnel Improvement Coordination (CAPES)
- National Council for Scientific and Technological Development (CNPq)
- Foundation to Support the Development of Education, Science and Technology of Mato Grosso do Sul State (FUNDECT)
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.
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