Identification of new Corynebacterium pseudotuberculosis antigens by immunoscreening of gene expression library

BMC Microbiology

Stenio Perdigão Fragoso¹  Carina Elisei de Oliveira²  Cleber Oliveira Soares³  Grácia Maria Soares Rosinha³  Cleber Eduardo Galvão⁴  Renata Ribeiro Bastos Pereira⁴  Odinéia Forner⁵ 

[1] Carlos Chagas Institute, Oswaldo Cruz Foundation (ICC-FIOCRUZ);Dom Bosco Catholic University (UCDB);Embrapa Beef Cattle, Animal Health - Animal Genetic Engineering Laboratory;Federal University of Mato Grosso do Sul (UFMS);Federal University of Paraná (UFPR);

关键词: Corynebacterium pseudotuberculosis;    Caseous lymphadenitis;    Sheep;    Gene expression;    Library;   

DOI  :  10.1186/s12866-017-1110-7

来源: DOAJ

【 摘 要 】

Abstract 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.

【 授权许可】

Unknown