Expression of functional recombinant human growth hormone in transgenic soybean seeds

• Published in: Transgenic research Vol. 20; no. 4; pp. 811 - 826
• Main Authors: Cunha, Nicolau B., Murad, André M., Cipriano, Thaís M., Araújo, Ana Cláudia G., Aragão, Francisco J. L., Leite, Adilson, Vianna, Giovanni R., McPhee, Timothy R., Souza, Gustavo H. M. F., Waters, Michael J., Rech, Elíbio L.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.08.2011; Springer; Springer Nature B.V
• Subjects: Amino Acid Sequence; Animal Genetics and Genomics; Antigens, Plant - genetics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; Coix lacryma-jobi; Fundamental and applied biological sciences. Psychology; Genetic Engineering; Genetic technics; Globulins - genetics; Glycine max; Glycine max - genetics; Glycine max - metabolism; Growth hormone; Human Growth Hormone - biosynthesis; Human Growth Hormone - genetics; Humans; Immunocytochemistry; Life Sciences; Mass spectroscopy; Methods. Procedures. Technologies; Molecular Medicine; Molecular Sequence Data; Original Paper; Plant Genetics and Genomics; Plant Proteins - genetics; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Promoter Regions, Genetic; Promoters; Protein Sorting Signals - genetics; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Regulatory sequences; Reproduction; Seed Storage Proteins - genetics; Seeds; Seeds - genetics; Seeds - metabolism; Signal peptides; Soybean Proteins - genetics; Soybeans; Transgenic animals and transgenic plants; Transgenics; Vacuoles; Vacuoles - metabolism; Transgenic; Molecular farming; Soybean seeds; Protein storage vacuoles; Human growth hormone; Human; Soybean; Vacuole; Protein; Somatotropin; Storage; Transgenic plant; Genetically modified organism; Transgenesis
• ISSN: 0962-8819; 1573-9368; 1573-9368
• DOI: 10.1007/s11248-010-9460-z

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.