4.2 Article

Expression of functional recombinant human growth hormone in transgenic soybean seeds

Journal

TRANSGENIC RESEARCH
Volume 20, Issue 4, Pages 811-826

Publisher

SPRINGER
DOI: 10.1007/s11248-010-9460-z

Keywords

Protein storage vacuoles; Soybean seeds; Transgenic; Molecular farming; Human growth hormone

Funding

  1. Empresa Brasileira de Pesquisa Agropecuaria (Embrapa)
  2. Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)

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We produced human growth hormone (hGH), a protein that stimulates growth and cell reproduction, in genetically engineered soybean [Glycine max (L.) Merrill] seeds. Utilising the alpha prime (alpha') subunit of beta-conglycinin tissue-specific promoter from soybean and the alpha-Coixin signal peptide from Coix lacryma-jobi, we obtained transgenic soybean lines that expressed the mature form of hGH in their seeds. Expression levels of bioactive hGH up to 2.9% of the total soluble seed protein content (corresponding to approximately 9 g kg(-1)) were measured in mature dry soybean seeds. The results of ultrastructural immunocytochemistry assays indicated that the recombinant hGH in seed cotyledonary cells was efficiently directed to protein storage vacuoles. Specific bioassays demonstrated that the hGH expressed in the soybean seeds was fully active. The recombinant hGH protein sequence was confirmed by mass spectrometry characterisation. These results demonstrate that the utilisation of tissue-specific regulatory sequences is an attractive and viable option for achieving high-yield production of recombinant proteins in stable transgenic soybean seeds.

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