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转基因大豆种子中功能性重组人生长激素的表达。

Expression of functional recombinant human growth hormone in transgenic soybean seeds.

机构信息

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

出版信息

Transgenic Res. 2011 Aug;20(4):811-26. doi: 10.1007/s11248-010-9460-z. Epub 2010 Nov 11.

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.

摘要

我们利用基因工程大豆[ Glycine max (L.)Merrill]种子生产了人生长激素(hGH),这是一种刺激生长和细胞繁殖的蛋白质。我们利用大豆β-伴大豆球蛋白组织特异性启动子的α'亚基和 Coix lacryma-jobi 的α- Coixin 信号肽,获得了在种子中表达 hGH 成熟形式的转基因大豆品系。在成熟的干燥大豆种子中,生物活性 hGH 的表达水平高达总可溶性种子蛋白含量的 2.9%(约 9 g kg(-1))。超微结构免疫细胞化学分析的结果表明,种子子叶细胞中的重组 hGH 被有效地定向到蛋白质储存液泡中。特异性生物测定表明,在大豆种子中表达的 hGH 具有完全的活性。通过质谱分析对重组 hGH 蛋白序列进行了确认。这些结果表明,利用组织特异性调控序列是在稳定的转基因大豆种子中实现重组蛋白高产的一种有吸引力和可行的选择。

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