An anti-amoebic vaccine: generation of the recombinant antigen LC3 from Entamoeba histolytica linked to mutated exotoxin A (PEΔIII) via the Pichia pastoris system.
Biotechnol Lett
; 39(8): 1149-1157, 2017 Aug.
Article
em En
| MEDLINE
| ID: mdl-28470625
ABSTRACT
OBJECTIVE:
To generate an immunogenic chimeric protein containing the Entamoeba histolytica LC3 fragment fused to the retrograde delivery domains of exotoxin A of Pseudomonas aeruginosa and KDEL3 for use as an effective vaccine.RESULTS:
A codon-optimized synthetic gene encoding the PEΔIII-LC3-KDEL3 fusion construct was designed for expression in Pichia pastoris. This transgene was subcloned into the plasmid pPIC9 for methanol-inducible expression. After transformation and selection of positive-transformed clones by PCR, the expression of the recombinant protein PEΔIII-LC3-KDEL3 was elicited. SDS-PAGE, protein glycosylation staining and western blot assays demonstrated a 67 kDa protein in the medium culture supernatant. The recombinant protein was detected with a polyclonal anti-6X His tag antibody and a polyclonal E. histolytica-specific antibody. A specific antibody response was induced in hamsters after immunization with this protein.CONCLUSIONS:
We report for the first time the design and expression of the recombinant E. histolytica LC3 protein fused to PEΔIII and KDEL3, with potential application as an immunogen.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Toxinas Bacterianas
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Proteínas Recombinantes de Fusão
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Vacinas
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ADP Ribose Transferases
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Fatores de Virulência
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Entamoeba histolytica
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Exotoxinas
Limite:
Animals
Idioma:
En
Revista:
Biotechnol Lett
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
México