Characterisation of alternative expression vectors for recombinant Bacillus Calmette-Guérin as live bacterial delivery systems



Título del documento: Characterisation of alternative expression vectors for recombinant Bacillus Calmette-Guérin as live bacterial delivery systems
Revista: Memorias do Instituto Oswaldo Cruz
Base de datos: PERIÓDICA
Número de sistema: 000437127
ISSN: 0074-0276
Autores: 1
2
1
Instituciones: 1Instituto Butantan, Laboratorio Especial de Desenvolvimento de Vacinas, Sao Paulo. Brasil
2Universidade Federal da Bahia, Faculdade de Farmacia, Salvador, Bahia. Brasil
Año:
Volumen: 115
País: Brasil
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Experimental, aplicado
Resumen en inglés Bacillus Calmette-Guérin (BCG) is considered a promising live bacterial delivery system. However, several proposals for rBCG vaccines have not progressed, mainly due to the limitations of the available expression systems. To obtain a set of mycobacterial vectors using a range of promoters with different strengths based on a standard backbone, previously shown to be stable. Mycobacterial expression vectors based on the pLA71 vector as backbone, were obtained inserting different promoters (PAN, PαAg, PHsp60, PBlaF* and PL5) and the green fluorescence protein (GFP) as reporter gene, to evaluate features such as their relative strengths, and the in vitro (inside macrophages) and in vivo stability. The relative fluorescence observed with the different vectors showed increasing strength of the promoters: PAN was the weakest in both Mycobacterium smegmatis and BCG and PBlaF* was higher than PHsp60 in BCG. The relative fluorescence observed in a macrophage cell line showed that PBlaF* and PHsp60 were comparable. It was not possible to obtain strains transformed with the extrachromosomal expression vector containing the PL5 in either species. We have obtained a set of potentially stable mycobacterial vectors with a arrange of expression levels, to be used in the development of rBCG vaccines
Disciplinas: Medicina
Palabras clave: Inmunología,
Bacterias,
BCG,
Vectores de expresión,
Recombinación genética,
Micobacterias,
Vacunas
Keyword: Immunology,
Bacteria,
Genetic recombination,
BCG,
Expression vectors,
Mycobacteria,
Vaccines
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