Large-scale production of foreign proteins via the novel plant transient expression system in Pisum sativum L.

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• 作者：Liang Li ; Xiufeng Wang ; Liping Yang ; Yajun Fan ; Xiaojuan Zhu…
• 刊名：Plant Biotechnology Reports
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：10
• 期：4
• 页码：207-217
• 全文大小：1,937 KB
• 刊物主题：Plant Sciences; Cell Biology; Plant Biochemistry; Biotechnology; Agriculture;
• 出版者：Springer Japan
• ISSN：1863-5474
• 卷排序：10

文摘

Transient expression of foreign genes by Agrobacterium infiltration is a versatile technique that can be used as a rapid tool for functional protein production in plants. A reproducible protocol of large-scale production of foreign proteins via the novel plant transient expression system in Pisum sativum L. was established in our study. Non-detached plants from soil-independent culture were used as the target organ, and vacuum infiltrating mediated by Agrobacterium tumefaciens harboring green fluorescent protein (GFP) gene was performed. Step-by-step optimization was performed and showed that the quality of plant material as well as agro-infiltration conditions were the major factors influencing the gene expression. Monitoring the transient GFP expression daily, the highest expression level was achieved on the 8th day post-infiltration. Evidence of anti-acidic fibroblast growth factor-single chain variable fragment (anti-aFGF-scFv) gene expression in pea seedling was also achieved using agro-mediated vacuum infiltration system. Our work proves that the system is suitable for the largescale production of pharmaceutical proteins. The in planta infiltration system described here provides a powerful tool to explore easily gene expression in Pisum sativum L. avoiding tissue culture steps and the labor-intensive generation of transgenic plants.KeywordsPisum sativum L.Transient expressionVacuum infiltrationGFPAnti-aFGF-scFv