High-Efficiency Expression and Production System of Heterologous Protein in Transgenic Rice Cell Cultur

Project: National Science and Technology CouncilNational Science and Technology Council Academic Grants

Project Details

Abstract

Molecular farming of valuable heterogonous proteins by plants has potential to product and create unlimited recombinant proteins. Plants and plant cells are now considered as viable and competitive expression systems for large-scale protein production. Stable transgenic plants could also produce leaves or seeds with rich recombinant protein for the purpose of long-term storage or direct processing. The disadvantages are farmland needed, commercial gene flow problem infield, and time-consuming of transgenic plants generated. By contrast, plant-suspension cells system can be used to produce recombinant proteins in large scale with high efficiency by fermentation. Therefore, the research and development of scale-up production technology of high-value proteins、vaccine or antibody by plant-suspension cells system is the most important issue in the whole biotechnology fields in these days. There are many advantages of immobilized system in biomaterial production. First, immobilized plant cell cultures is better than freely suspended cells in both the practical bioreactor configuration and the physiological state of the cells during the culture period. Second, immobilized system can increase production by very high degree cell aggregation in system, because high cell aggregation could increase better cell-cell contact and cause better cell differentiation by reducing cell division rate. Third, immobilization also offers advantages from a process engineering point of view, especially in providing an easily manipulated, low shear environment. In this study, we will develop a simple and rapid technology for high efficiency heterologous protein production in suspension-cultured transgenic rice cells. Here, five keystone technology will be developed:(1)A whole new design of rice suspension cell expression cassette for multiple variable proteins production;(2)A simple and rapid transformation system;(3)A application of secret signal peptide;(4)the development of novel GFP production indicator which could simplify the monitor analysis operation during the developmental downstream process of this technology;(5)A scale-up immobilization fermentation process for efficient target proteins production. In order to investigate the feasibility of the heterologous protein production technology platform, the cDNA of recombination granulocyte-macrophage colony stimulating factor (GM-CSF) will be chosen as a target product in this study. In addition, the optimal production condition and operation model will be investigated by the technology platform. It could fast and completely carry out the whole production process from cDNA fragment to the target proteins with high efficiency. Moreover, the equipment and cost should be reduced as low as possible. By combination and coherence of different high technology with higher efficiency and lower cost, we can predict the tremendous potential of this heterologous protein production technology platform.

Project IDs

Project ID:PD9408-0426
External Project ID:NSC94-2317-B182-001
StatusFinished
Effective start/end date01/08/0531/12/06

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