Expression of functional recombinant human growth hormone in transgenic soybean seeds
Expression of functional recombinant human growth hormone in transgenic soybean seeds
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DOI：
10.1007/s11248-010-9460-z
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发表时间：
2011-08-01
影响因子：
3
通讯作者：
Rech, Elibio L.
中科院分区：
文献类型：
--
作者：
Cunha, Nicolau B.;Murad, Andre M.;Rech, Elibio L.
我们在基因工程大豆[Glycine max（L.）梅里尔]种子。利用来自大豆的β-伴大豆球蛋白组织特异性启动子的α '亚基和来自薏苡的α-薏苡蛋白信号肽，我们获得了在其种子中表达成熟形式hGH的转基因大豆品系。在成熟的干大豆种子中测得生物活性hGH的表达水平高达总可溶性种子蛋白含量的2.9%（相当于约9 g kg（-1））。超微结构的免疫细胞化学分析结果表明，重组hGH在种子子叶细胞中被有效地导向蛋白质储存液泡。特异性生物测定表明，在大豆种子中表达的hGH是完全活性的。通过质谱表征确认重组hGH蛋白序列。这些结果表明，利用组织特异性调节序列是在稳定的转基因大豆种子中实现重组蛋白高产率的一个有吸引力且可行的选择。
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.