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Doctoral Thesis
DOI
10.11606/T.87.2013.tde-14052014-095103
Document
Author
Full name
Alex Issamu Kanno
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Leite, Luciana Cezar de Cerqueira (President)
Barbuto, Jose Alexandre Marzagao
Leão, Sylvia Luisa Pincherle Cardoso
Maldonado, Gabriel Padilla
Marco, Ricardo De
Title in Portuguese
Expressão de antígenos de Schistosoma mansoni em BCG recombinante.
Keywords in Portuguese
Mycobacterium bovis
Schistosoma mansoni
Mutagênese
Vacinas
Abstract in Portuguese
O BCG é um bacilo atenuado de Mycobacterium bovis atualmente investigado na expressão de antígenos heterólogos. Genes oriundos de vírus, bactérias e parasitas são clonados em vetores de expressão micobacterianos e a partir deles, são geradas cepas de BCG recombinante, rBCG. Duas cepas micobacterianas, BCG e M. smegmatis, demonstraram a expressão do antígeno SmStoLP-2 e animais imunizados com este BCG não demonstra diferença na proteção contra a infecção por cercárias, apesar da resposta imune diferenciada. A otimização do códon de SmTSP-2 e SmVAL-5 permitiu sua expressão em M. smegmatis recombinante, mas não em BCG. M. smegmatis transformado com uma biblioteca de plasmídeos contendo o promotor L5p mutagenizado à montante de egfp demonstra acentuada diferença na fluorescência. 3 destes plasmídeos definidos como ''forte'' e ''fraco'', com base em sua capacidade de produzir GFP foram utilizados para expressar em BCG o antígeno Sm29 em maior e menor nível.
Title in English
Expression of Schistosoma mansoni antigens in recombinant BCG.
Keywords in English
Mycobacterium bovis
Schistosoma mansoni
Mutagenesis
Vaccine
Abstract in English
BCG is an attenuated bacillus derived from Mycobacterium bovis currently being investigated to express foreign antigens. Genes from viruses, bacteria and parasites are cloned to mycobacterial vectors and with them, recombinant BCG strains are created. Recombinant BCG and M. smegmatis strains where capable to express SmStoLP-2 and mice immunized with these rBCGs do not show better protection against infection with cercariae, although the distinct immune response observed. The use of codon optimization made possible the expression of SmTSP-2 and SmVAL-5 in M. smegmatis but not BCG. M. smegmatis transformed with a library of plasmids containing the L5p upstream egfp gene show a wide range in fluorescence. 3 of these plasmids strong and weak defined as their ability to produce GFP were used to express in BCG the antigen Sm29 at different levels by each promoter.
 
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Publishing Date
2014-05-21
 
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