State Key Laboratory of Genetic Engineering, School of Life Sciences, Morgan-Tan International Center for Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Fudan University, Shanghai, People's Republic of China.
Biotechnol Appl Biochem. 2011 May;58(3):139-46. doi: 10.1002/bab.30. Epub 2011 Jun 5.
Novel protein expression in plant-based systems has become an important tool in producing and studying therapeutic proteins. Among many plant-based systems developed so far, oleosin fusion technology is one of the most cost-effective and convenient methods. In this study, an important therapeutic protein, human insulin-like growth factor 1 (hIGF-1), was expressed in Arabidopsis thaliana seeds via this technology. The plant bias codon usage-optimized hIGF-1 gene was fused to the C-terminal of A. thaliana 18.5 kDa oleosin gene, and the fusion gene driven by an oleosin promoter was transferred into A. thaliana ecotype Col-0. The accumulation of oleosin-hIGF-1 fusion protein in transgenic seeds was up to 0.75% of total seed protein (TSP) and the expression level of hIGF-1 was 0.17% of the TSP, which was eight times higher than previously reported using other plant-based hIGF-1 production systems. The biological activity of the hIGF-1 as an oleosin-hIGF-1 fusion protein in vitro was demonstrated by using human SH-SY5Y neuroblastoma cells.
在植物生物系统中表达新型蛋白质已成为生产和研究治疗性蛋白质的重要工具。在迄今为止开发的许多植物生物系统中，油体融合技术是最具成本效益和便利的方法之一。在这项研究中，通过该技术在拟南芥种子中表达了一种重要的治疗性蛋白质，人胰岛素样生长因子 1（hIGF-1）。将经过植物偏爱密码子使用优化的 hIGF-1 基因融合到拟南芥 18.5 kDa 油体蛋白基因的 C 端，并用油体启动子驱动融合基因转入拟南芥生态型 Col-0。油体-hIGF-1 融合蛋白在转基因种子中的积累量高达总种子蛋白（TSP）的 0.75%，hIGF-1 的表达量为 TSP 的 0.17%，比以前使用其他植物 hIGF-1 生产系统报道的表达水平高 8 倍。通过体外使用人 SH-SY5Y 神经母细胞瘤细胞，证明了 hIGF-1 作为油体-hIGF-1 融合蛋白的生物活性。
