Purification, Characterization, and Molecular Cloning of A Viral Protease Inhibitor

病毒蛋白酶抑制剂的纯化、表征和分子克隆

• 批准号: 9100036
• 负责人: Chao-yun Shih
• 金额: $ 36.16万
• 依托单位: Southern University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-15 至 1995-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9100036&HistoricalAwards=false
• 关键词: Purification; Characterization; Molecular; Cloning; Viral

Proteolytic cleavage of precursor proteins by virus coded proteases is a common protein synthesis strategy utilized by many animal and plant viruses for synthesis of functional viral proteins. Prevention of these cleavage processes may offer a specific means for controlling replication of these viruses. The proposed project is designed to investigate a protease inhibitor which Southern University biologists found to exist in cowpea embryos. This inhibitor can completely prevent proteolytic processing of the precursor proteins of cowpea mosaic virus (CPMV) in an in vitro protein synthesizing system. The plan is to develop a purification procedure for complete purification of the inhibitor, to characterize the inhibitor biochemically, to compare the activity of the inhibitor to the activities of synthetic peptides, to determine the existence or absence of this inhibitor in CPMV resistance and sensitive cowpea plants, and to determine the amino acid sequence of the inhibitor using DNA cloning and sequencing methods. The long range goals of this research are to gain a complete understanding of the structural and functional properties of this inhibitor, and to use this inhibitor as a model for examining the possibility of utilizing protease inhibitors as antiviral agents for prevention or treatment of viral diseases. Besides the research goals, an additional objective of this proposed project is to present an opportunity to minority students and faculty of Southern University and other minority institutions to receive research training and to gain experience in modern molecular biology techniques, including genetic engineering techniques. This objective will be accomplished through seminars and workshops.

病毒编码的前体蛋白的蛋白水解切割 蛋白酶是一种常见的蛋白质合成策略， 许多动物和植物病毒的合成功能 病毒蛋白 防止这些裂解过程可 提供了一种特定的方法来控制这些复制 病毒 该项目旨在研究一种蛋白酶 南方大学的生物学家发现， 存在于豇豆胚胎中。 这种抑制剂可以完全 防止前体蛋白的蛋白水解加工， 豇豆花叶病毒（CPMV）的体外蛋白质 合成系统 计划是开发一种净化 完全纯化抑制剂的程序， 对抑制剂进行生物化学表征， 抑制剂对合成酶活性的抑制作用 肽，以确定是否存在这种 抑制剂在CPMV抗性和敏感豇豆植物中， 并确定抑制剂的氨基酸序列 使用DNA克隆和测序方法。 远程 本研究的目的是全面了解 它的结构和功能特性 抑制剂，并使用这种抑制剂作为模型， 研究利用蛋白酶抑制剂 作为抗病毒剂用于预防或治疗病毒感染 疾病 除了研究目标之外，本研究的另一个目标是 该项目旨在为少数民族提供一个机会， 南方大学的学生和教师以及其他 少数群体机构接受研究培训， 获得现代分子生物学技术的经验， 包括基因工程技术。 这一目标 将通过研讨会和讲习班完成。