Banca de DEFESA: JÚLIO CÉSAR BRAGA DE SOUZA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : JÚLIO CÉSAR BRAGA DE SOUZA
DATE: 12/05/2023
TIME: 14:00
LOCAL: https://meet.google.com/asg-dpnh-fgn
TITLE:

Analysis and selection of antigenic proteins from Mycoplasma bovis in the search for vaccine candidates or diagnosis

KEY WORDS:

M. bovis; immunodiagnosis; bovine infections; recombinant protein; reverse vaccinology.

PAGES: 90
BIG AREA: Ciências Biológicas
AREA: Microbiologia
SUBÁREA: Microbiologia Aplicada
SPECIALTY: Microbiologia Médica
SUMMARY:

Mycoplasma bovis is one of the main bovine pathogens and is present in the etiology of mastitis, respiratory diseases, and arthritis. Treatment of these diseases is limited. Thus, prevention proves to be an important alternative. However, in Brazil, there is no data on effective vaccines against M. bovis. Therefore, this study aimed to analyze and select conserved proteins in M. bovis for validation and application in vaccines and immunodiagnostic tests. The targets were set using bioinformatics tools, using the UniProt database to obtain the proteomes. Proteins were evaluated using PsortB for subcellular localization. The following predictions were realized: the presence of transmembrane regions and signal peptides using the TopCons database. Homology between the two M. bovis proteomes was also evaluated using the public CD-HIT program. For epitope prediction, the consensus of tools provided by IEDB was used to predict B cell epitopes. NETMHCcons and NETMHCIIpan 3.1 were used to predict T cell epitopes presented in MHC I and MHC II pathways, respectively. The selected proteins were then cloned and expressed in Escherichia coli cells. After purification, the proteins were evaluated for antigenicity and validated using dot blotting assays with rabbit serum known to be contaminated with M. bovis. The protein A0A0Y59Y3U4 was predicted by the consensus of bioinformatics tools as a good target for study. Optimization of its expression was carried out using six strains of E. coli cells, and four of them (Arctic, pLysS, BL21, and C41) showed significant results, with C41 showing the best yield. After purification, the protein was evaluated for reactivity against sera known to be infected with M. bovis, and the protein reacted to the tested sera. Thus, the results obtained in the present study open up possibilities to advance studies using this protein in stages of animal experimentation in search of an effective vaccine candidate to control this disease.

COMMITTEE MEMBERS:
Presidente - 1561874 - LUCAS MIRANDA MARQUES
Externo à Instituição - MANOEL NERES SANTOS JUNIOR - UFBA
Externo à Instituição - MAYSA SANTOS BARBOSA