Seleção, caracterização e clonagem dos genes fljB e groEL agonistas dos receptores de reconhecimento de padrão do sistema imune inato das aves

Pesquisa Veterinária Brasileira

Bruno A. Soares²  Solange A. Ságio¹  Ana P. Peconick¹  Priscilla R. Barrios¹  Antônio Chalfun-júnior¹  Geraldo M. Costa¹  Joziana M.p. Barçante¹  Nelson R.s. Martins² 

[1] ,Universidade Federal de Minas Gerais Escola de Veterinária Laboratório de Doenças das AvesBelo Horizonte MG ,Brasil

关键词: Recombinant protein;    PAMP;    flagellin;    DAMP;    HSP60;    birds;    Proteína recombinante;    PAMP;    flagelina;    DAMP;    HSP60;    aves;   

DOI  :  10.1590/S0100-736X2014000300004

来源: SciELO

【 摘 要 】

The recombinant production of innate immune system pattern recognition receptor agonists has provided a new tool for the production of immunostimulants for animals. The molecular pattern associated with the pathogen (PAMP), flagellin, coded by the fljB gene from Salmonella Typhimirium, and the molecular pattern associated to the damage (DAMP), HSP60, coded by the groEL gene from S. Typhimurium and S. Enteritidis, are recognized by pattern recognition receptors (PRRs) of the innate immune system of birds. In the present study, we performed the cloning of genetic fragments of the genes fljB, from S. Typhimurium, and groEL from S. Typhimurium and S. Enteritidis inserted in expression vector pET100/D-TOPO and transformed in E. coli TO10 cells. The clones were evaluated by colony PCR, plasmidial DNA PCR and genome sequencing in order to confirm the presence of these genes. In the colony PCR, we identified the presence of genes groEL (S. Enteritidis), groEL (S. Typhimurium) and fljB (S. Typhimurium) in 80%, 60% and 80% of the transformed colonies, respectively. The cloning system adopted allowed the production of HSP60 genetic fragment clones and flagellin of Salmonella strains, allowing the posterior use of these clones in gene expression trials, with the future potential of being used as non-specific immunostimulants for birds.

【 授权许可】

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