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Expression of functional recombinant human growth hormone in transgenic soybean seeds

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Abstract

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 (α′) subunit of β-conglycinin tissue-specific promoter from soybean and the α-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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Acknowledgements

The authors are grateful to Warley Almeida and Luis Carlos Lemos for technical assistance and to Ana C.M.M. Gomes for transmission electron microscopy assistance. The authors thank Prof. Roger N. Beachy (Danforth Center, USA) for providing the α′ subunit of β-conglycinin regulatory sequences.

This study was supported in part by the Empresa Brasileira de Pesquisa Agropecuária (Embrapa) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Authors and Affiliations

  1. Embrapa Genetic Resources and Biotechnology, Parque Estação Biológica (PqEB), Av. W5 Norte, Brasília, DF, 70770-917, Brazil

    Nicolau B. Cunha, André M. Murad, Thaís M. Cipriano, Ana Cláudia G. Araújo, Francisco J. L. Aragão, Giovanni R. Vianna & Elíbio L. Rech

  2. Department of Cellular Biology, University of Brasília, Brasília, DF, 70910-900, Brazil

    Nicolau B. Cunha

  3. Center of Molecular Biology and Genetic Engineering (CBMEG), University of Campinas, Sao Paulo, SP, 13083-875, Brazil

    Adilson Leite

  4. Institute for Molecular Bioscience, University of Queensland, St Lucia, QLD, Australia, 4072

    Timothy R. McPhee & Michael J. Waters

  5. MS Applications Research and Development Laboratory, Waters Corporation, Alameda Tocantins 125, 27th Floor, West Side, Alphaville, São Paulo, SP, 06455-020, Brazil

    Gustavo H. M. F. Souza

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  1. Nicolau B. Cunha
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  2. André M. Murad
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Correspondence to Elíbio L. Rech.

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In memoriam of Adilson Leite.

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Cunha, N.B., Murad, A.M., Cipriano, T.M. et al. Expression of functional recombinant human growth hormone in transgenic soybean seeds. Transgenic Res 20, 811–826 (2011). https://doi.org/10.1007/s11248-010-9460-z

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