Extended DNA amplification with thermostable chaperons

使用热稳定性伴侣扩展 DNA 扩增

• 批准号: 6882777
• 负责人: Frank Thomson Robb
• 金额: $ 14.34万
• 依托单位: FIDELITY SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882777
• 关键词: DNA; DNA directed DNA polymerase; DNA topoisomerases; RNA directed DNA polymerase; biotechnology; chemical reaction; enzyme activity; enzyme structure; high throughput technology; ligase; molecular chaperones; nucleic acid amplification techniques; nucleic acid denaturation; nucleic acid sequence; polymerase chain reaction; protein engineering; protein folding; synthetic enzyme; thermostability

DESCRIPTION (provided by applicant): The goal of this project is to develop a general method of improving the stability and durability of enzymes used in DNA and RNA amplification, sequencing, restriction analysis and ligation. The enzymes to be improved include DNA polymerases, topoisomerases, reverse transcriptases and ligases. In the event this goal can be achieved, broad impacts may result in the areas of clinical, forensic and basic science that rely on DNA sequencing, amplification and RT-PCR processes. To do this, extrinsic stabilization and refolding of proteins mediated by chaperones from hyperthermophiles will be adapted for incorporation into reaction mixtures with DNA modifying enzymes.

描述（由申请人提供）：本项目的目标是开发一种提高DNA和RNA扩增、测序、限制性分析和连接中所用酶的稳定性和耐久性的通用方法。待改进的酶包括DNA聚合酶、拓扑异构酶、逆转录酶和连接酶。如果这一目标能够实现，可能会对依赖DNA测序、扩增和RT-PCR过程的临床、法医和基础科学领域产生广泛影响。要做到这一点，外在的稳定和重折叠的蛋白质介导的分子伴侣从超嗜热菌将适用于纳入反应混合物与DNA修饰酶。

项目成果

Stabilization of Taq DNA polymerase at high temperature by protein folding pathways from a hyperthermophilic archaeon, Pyrococcus furiosus.

• DOI: 10.1002/bit.20781
• 发表时间: 2006-01
• 期刊: Biotechnology and bioengineering
• 影响因子: 3.8
• 作者: P. Laksanalamai;A. Pavlov;A. Slesarev;F. Robb