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Publicação
Culturas Celulares de Tabaco BY2 para a Produção de Biofármacos: O Papel da Metilação do ADN na Expressão do Transgene
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| 3.92 MB | Adobe PDF |
Orientador(es)
Resumo(s)
"Genetic engineering strategies such as molecular farming, essential for the production of recombinant
proteins and other secondary metabolites in plants, have the potential to provide virtually unlimited
amounts of recombinant proteins for use as diagnostic and therapeutic tools in health care and life
sciences. Tobacco cell suspension cultures such as BY2 (Bright Yellow 2) can be easily transformed
and a number of approaches have been developed to increase production, including medium
optimization or the use of improved expression vectors. The aim of this work was to try to optimize the
production, by increasing the transcription levels of the transgene encoding the protein of interest
lipocalin-type Prostaglandin D synthase (L-PGDS), through epigenetic modulation using
hypomethylation agents. Treatment with 5-azacytidine in plant and animal cells results in demethylation
of DNA both directly and indirectly. We attempted to establish the optimum concentration of 5-
azacytidine in the cell lines by applying different concentrations of the compound, and subsequently
performed the immunodetection of methylated cytosines in BY2 nuclei, as well as methylation studies
using Dot-Blot methods and conversion by bisulfite.(...)
Descrição
Palavras-chave
Molecular Farming Nicotiana tabacum Bright Yellow 2 (BY2) 5-azacitidina