ExpressÃo de cinco diferentes proteÃnas de interesse farmacolÃgico em sementes transgÃnicas de soja [Glycine Max L. (Merril)]

AUTOR(ES)
DATA DE PUBLICAÇÃO

2008

RESUMO

Superior plants represent a convenient system for large-scale production of heterologous proteins. Its potential has been confirmed by the commercial success of such production systems, based on genetic transformation of a broad range of vegetable species, resulting in efficient production of more than 100 different heterologous proteins. A representative percentage of recombinant polypeptides synthesized in transgenic plants is based in molecules with pharmacological applications, such as hormones, growth factors, antibodies and coagulation factors. Plant-based bioreactors presents a series of advantages when compared with other production systems, specially in what concerns of economy, easy-hand manipulation and biosafety, besides some homogeneity, yeld-production and stability limitations. The choice for Soybean plants [Glycine max L.(Merril)] for function like expression vehicles of biopharmaceutical proteins means not only to solve such problems but to explore natural and favorable abilities presented by these organisms such as low allogamy frequency, natural absence of related plants in Brazil and its excellence in seed protein accumulation. Tissue-specific expression and protein targeting of heterologous proteins to protein bodies can maximize their accumulatin in transgenic seeds. Different expression cassettes were constructed in our laboratory, with the purpose to express five different coding sequences of social impact proteins such as the human pro-insulin, growth hormone and coagulation factor IX, and two recombinant antibodies: sc FvDIR83D4 and anti- CD18, under control of the tissue-specific β-Phaseolin promoter (the most abundant In bean seeds) and α-coixin signal-peptide (subcellular cotiledonar target sequence). After gene cloning, different soybean transgenic lines were obtained by biobalistics, and molecular and biochemical analysis were carried out to determine the copy number of transgenic sequences integrated in plant genome, transgene segregation in alternate generations, protein accumulation levels, gene expression kinetics during plant cycle and subcellular targeting.

ASSUNTO(S)

anticorpos recombinantes fator de coagulaÃÃo sangÃÃnea human growth hormone hormÃnio de crescimento humano soja, soja â melhoramento genÃtico; insulina; hormÃnios plant-based bioreactors insulina humana transgenic soybean soja transgÃnica biorreatores vegetais human insulin coagulation factor ciÃncias biolÃgicas proteÃnas heterÃlogas heterologous proteins recombinant antibodies

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