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Master's Dissertation
DOI
10.11606/D.42.2017.tde-20022017-144035
Document
Author
Full name
Maysa Santos Barbosa
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Timenetsky, Jorge (President)
Fernandes, Andrea Balan
Gregory, Lilian
Lourenço, Alexandre
Title in Portuguese
Clonagem, purificação e caracterização de proteínas antigênicas recombinantes obtidas de Mycoplasma agalactiae.
Keywords in Portuguese
Mycoplasma agalactiae
Agalaxia contagiosa
Antígenos
Proteína recombinante
Abstract in Portuguese
A agalaxia contagiosa é uma doença de notificação obrigatória que causa severas perdas econômicas para produção de ovinos e caprinos mundialmente. Apesar de seu impacto na produção animal, pouco se sabe sobre os fatores de virulência e patogenicidade do M. agalactiae (principal agente etiológico). Desta maneira, o presente estudo possuiu como objetivo a identificação, purificação e caracterização de proteínas antigênicas de M. agalactiae. Para tanto, quatro proteínas de superfície com potencial antigênico (WP_011949419.1, WP_011949418.1 (P40), WP_011949336.1, WP_011949770.1) foram selecionadas. Essas proteínas foram expressas em Escherichia coli e purificadas em coluna de níquel. As proteínas purificadas foram avaliadas quanto a antigenicidade em Western blotting utilizando soros de caprinos naturalmente infectados com M. agalactiae. Todas as proteínas expressas foram imunorreativas aos soros de caprinos naturalmente infectados, demonstrando que as proteínas utilizadas nesse estudo são possivelmente antigênicas e possuem epítopos acessíveis.
Title in English
Cloning, purification and characterization of recombinant antigenic protein of Mycoplasma agalactiae.
Keywords in English
Mycoplasma agalactiae
Agalactia contagious
Antigens
Recombinant protein
Abstract in English
The Contagious agalactia is a notifiable disease that causes severe economic losses to sheep and goats worldwide. Despite its impact on animal production, little is known about the virulence factors and pathogenicity of M. agalactiae (main etiological agent). Thus, the present study identified, purified and characterized antigenic proteins of M. agalactiae. Therefore, four surface proteins with antigenic potential (WP_011949419.1, WP_011949418.1 (P40), WP_011949336.1, WP_011949770.1) were selected. These proteins were expressed in Escherichia coli and purified on nickel column. The purified proteins were assayed for antigenicity by Western blotting using goat sera naturally infected with M. agalactiae. All expressed proteins were immunoreactive with sera from naturally infected goats, demonstrating that the proteins used in this study are possibly antigenics and it have acessible epitopes.
 
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Release Date
2019-02-20
Publishing Date
2017-02-20
 
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