Collaborative Research: Transient Protein Expression in Plant Tissue Culture

合作研究：植物组织培养中的瞬时蛋白表达

• 批准号: 0003926
• 负责人: Wayne Curtis
• 金额: $ 22.67万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-10-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0003926&HistoricalAwards=false
• 关键词: Collaborative; Research; Transient; Protein; Expression

The Principal Investigators (PIs)' overall objective is to develop plant cell and tissue culture technology, which will permit rapid evaluation of heterologous gene expression in plants. This work includes the development of rigorous principles for the monitoring, control and operation of bioreactors for plant cell and tissue culture. These bioreactor operational principles will be utilized to correlate performance of transient expression systems using Agrobacterium co-cultivation for DNA transformation and the TMV vector or a single-stranded DNA geminivirus vector capable of DNA amplification for protein production (based on tobacco and corn seed endosperm) while testing expression and accumulation of three therapeutic proteins: a monoclonal antibody (NR-LU-10 MAb), a cytokine (human growth hormone, hGH), and a blood transport protein (human transferrin, hTR). The specific objectives are to: (1) Develop operational and control strategies for bioreactor growth of plant cell and root culture, (2) Develop and optimize techniques / conditions for DNA delivery for transient expression, and (3) Utilize bioreactor control strategies and optimized transient therapeutic protein expression systems to evaluate heterologous transgene expression in plant cell and tissue culture, followed by correlation with performance in transgenic plants with the same proteins. This is a collaborative GOALI project involving three PIs with different complementary scientific and technological expertise. Wayne Curtis the primary PI at the Pennsylvania State University is an expert in the area of bioreactor technology, Hugh Mason the second PI at the Boyce Thompson Institute and Cornell University in the area of transgenic plants, and Doug Russell at the Integrated Protein Technologies of Monsanto in the area of production of proteins from transgenic plants.

主要研究者（PI）的总体目标是开发植物细胞和组织培养技术，这将允许快速评估植物中异源基因的表达。这项工作包括为植物细胞和组织培养生物反应器的监测、控制和操作制定严格的原则。这些生物反应器的操作原理将用于关联瞬时表达系统的性能，所述瞬时表达系统使用农杆菌共培养进行DNA转化，并且使用TMV载体或能够进行DNA扩增以产生蛋白质的单链DNA双生病毒载体（基于烟草和玉米种子胚乳），同时测试三种治疗性蛋白质的表达和积累：单克隆抗体（NR-LU-10 MAb）、细胞因子（人生长激素，hGH）和血液转运蛋白（人转铁蛋白，hTR）。具体目标是：（1）开发用于植物细胞和根培养物的生物反应器生长的操作和控制策略，（2）开发和优化用于瞬时表达的DNA递送的技术/条件，和（3）利用生物反应器控制策略和优化的瞬时治疗性蛋白质表达系统来评估植物细胞和组织培养物中的异源转基因表达，随后是与具有相同蛋白质的转基因植物中的性能的相关性。 这是一个GOALI合作项目，涉及三个具有不同互补科学和技术专长的PI。宾夕法尼亚州立大学的主要PI韦恩柯蒂斯是生物反应器技术领域的专家，博伊斯汤普森研究所和康奈尔大学的第二PI休梅森在转基因植物领域，和道格罗素在孟山都的综合蛋白质技术领域的蛋白质生产转基因植物。