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浮萍作为药物替代生产系统的案例研究:抑肽酶

Spirodela (duckweed) as an alternative production system for pharmaceuticals: a case study, aprotinin.

作者信息

Rival Sandrine, Wisniewski Jean-Pierre, Langlais Audrey, Kaplan Hélène, Freyssinet Georges, Vancanneyt Guy, Vunsh Ron, Perl Avihai, Edelman Marvin

机构信息

LemnaGene SA, 71 Chemin du Moulin Carron, Dardilly, France.

出版信息

Transgenic Res. 2008 Aug;17(4):503-13. doi: 10.1007/s11248-007-9123-x. Epub 2007 Aug 10.

Abstract

Aprotinin is a small serine protease inhibitor used in human health. Spirodela were transformed, via Agrobacterium, with a synthetic gene encoding the mature aprotinin sequence and a signal peptide for secretion which was driven by the CaMV 35S promoter. A total of 25 transgenic Spirodela lines were generated and aprotinin production was confirmed by northern and western blot analyses. Expression levels of up to 3.7% of water soluble proteins were detected in the plant and 0.65 mg/l in the growth medium. In addition, immunoaffinity purification of the protein followed by amino acid sequencing confirmed the correct splicing of the aprotinin produced in Spirodela and secreted into the growth medium.

摘要

抑肽酶是一种用于人类健康的小型丝氨酸蛋白酶抑制剂。通过农杆菌介导,将编码成熟抑肽酶序列和用于分泌的信号肽的合成基因导入紫萍,该基因由花椰菜花叶病毒(CaMV)35S启动子驱动。共产生了25个转基因紫萍株系,通过Northern印迹和Western印迹分析证实了抑肽酶的产生。在植物中检测到水溶性蛋白的表达水平高达3.7%,在生长培养基中为0.65mg/L。此外,对该蛋白进行免疫亲和纯化后进行氨基酸测序,证实了紫萍中产生并分泌到生长培养基中的抑肽酶的正确剪接。

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