| Journal: | Memorias do Instituto Oswaldo Cruz |
| Database: | PERIÓDICA |
| System number: | 000437127 |
| ISSN: | 0074-0276 |
| Authors: | Nascimento, Larissa V1 Santos, Carina C2 Leite, Luciana C.C1 |
| Institutions: | 1Instituto Butantan, Laboratorio Especial de Desenvolvimento de Vacinas, Sao Paulo. Brasil 2Universidade Federal da Bahia, Faculdade de Farmacia, Salvador, Bahia. Brasil |
| Year: | 2020 |
| Volumen: | 115 |
| Country: | Brasil |
| Language: | Inglés |
| Document type: | Artículo |
| Approach: | Experimental, aplicado |
| English abstract | 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 |
| Disciplines: | Medicina |
| Keyword: | Inmunología, Bacterias, BCG, Vectores de expresión, Recombinación genética, Micobacterias, Vacunas |
| Keyword: | Immunology, Bacteria, Genetic recombination, BCG, Expression vectors, Mycobacteria, Vaccines |
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