GENE ENGINEERING AND COMBINATIONAL BIOLOGY

描述(调查人员摘要)：本文件中描述的研究计划 应用的重点是新基因的开发和实施 工程学方法。正在开发的方法特别是 适合于模组基因的操作，如多肽合成基因。 我们的目标是将模块化基因分解成它们的组成部分模块，并使用 这些模块作为组装新的模块基因的构建块。 为了实现这一目标，我们开发了两种新的基因工程方法。 这两种办法都将在供资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 创建嵌合的RNA-悬垂克隆(ROC)和NA-悬垂克隆(DOC) 基因。最后，核酶方法将与ROC和OC方法相结合 创造一种新的基因工程系统，充分利用 个别方法的长处。 有效的基因工程方法对于这两种基本的 以及对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被用来创造 编码嵌合肽合成酶基因的组合基因库。 已知自然产生的多肽合成酶可以合成重要的 抗生素，如青霉素和万古霉素。此外， 免疫抑制剂环孢素是由一种多肽合成酶产生的。这个 在这个项目过程中产生的嵌合多肽合成酶基因 应该编码生物合成新奇的杂交酶 活性分子。这个项目的一个长期目标是筛选嵌合基因。 用于识别合成新化合物的酶的文库可能是 发展成毒品。

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.