GENE ENGINEERING AND COMBINATIONAL BIOLOGY

基因工程和组合生物学

• 批准号: 6229828
• 负责人: KEVIN A JARRELL
• 金额: $ 31.03万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-15 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6229828
• 关键词: RNA splicing; biotechnology; enzyme biosynthesis; fusion gene; genetic library; genetic manipulation; genetic recombination; genetic techniques; mass spectrometry; molecular cloning; nucleic acid sequence; peptide library; polymerase chain reaction; ribozymes; technology /technique development

DESCRIPTION (Investigator's Abstract): The research program described in this application is focused on the development and implementation of new gene engineering methods. The methods that are being developed are particularly suited to the manipulation of modular genes, such as peptide synthetic genes. Our goal is to break the modular genes up into their component modules, and use those modules as building blocks for the assembly of novel new modular genes. To achieve this goal, we have developed two new approaches to gene engineering. Both approaches will be optimized and implemented during the funding period. The first approach involves the use of ribozymes(i.e., enzymes comprised of RNA) as tools for chimeric gene assembly. The second approach involves the use of RNA-overhang cloning (ROC) and NA-overhang cloning (DOC) to create chimeric genes. Finally, the ribozyme method will be combined with the ROC and OCmethods to create a new gene engineering system that takes full advantage of the strengths of the individual approaches. Efficient gene engineering methods are of fundamental importance for both basic and applied research on topics directly relevant to human health. For example, the gene engineering methods described here will be used to create combinatorial gene libraries that encode chimeric peptide synthetase genes. Naturally occurring peptide synthetases are known to synthesize important antibiotics, such as penicillin and vancomycin. Furthermore, the immunosuppressant cyclosporine is produced by a peptide synthetase. The chimeric peptide synthetase genes generated during the course of this project should encode hybrid enzymes that synthesize novel biologically active-molecules. A long-term goal of this project is to screen chimeric gene libraries to identify enzymes that synthesize novel compounds at could be developed as drugs.

描述（研究者摘要）：本文描述的研究计划 应用重点是新基因的开发和实施 工程方法。正在开发的方法特别是 适合操作模块化基因，例如肽合成基因。 我们的目标是将模块化基因分解为其组件模块，并使用 这些模块作为组装新模块基因的构建块。 为了实现这一目标，我们开发了两种新的基因工程方法。 这两种方法都将在资助期间得到优化和实施。 第一种方法涉及使用核酶（即由以下组成的酶） RNA）作为嵌合基因组装的工具。第二种方法涉及使用 RNA 突出克隆 (ROC) 和 NA 突出克隆 (DOC) 来创建嵌合体 基因。最后，核酶法将与ROC和OC法相结合 创建一个新的基因工程系统，充分利用 个人方法的优势。 高效的基因工程方法对于基础和基础研究都至关重要。 以及与人类健康直接相关的主题的应用研究。例如， 这里描述的基因工程方法将用于创建 编码嵌合肽合成酶基因的组合基因文库。 已知天然存在的肽合成酶可以合成重要的肽 抗生素，例如青霉素和万古霉素。此外， 免疫抑制剂环孢菌素由肽合成酶产生。这 在该项目过程中产生的嵌合肽合成酶基因 应该编码合成新生物的杂合酶 活性分子。该项目的长期目标是筛选嵌合基因 识别合成新化合物的酶的文库可能是 被开发为药物。