Funktionelle Genomik mit mRNA-Protein-Konjugaten und DNA Mikrochips

利用 mRNA-蛋白质缀合物和 DNA 微芯片进行功能基因组学

• 批准号: 5251680
• 负责人: Professor Dr. Burckhard Seelig
• 金额: --
• 依托单位: BioTechnology Institute
• 依托单位国家: 德国
• 项目类别: Emmy Noether International Fellowships
• 财政年份: 2000
• 资助国家: 德国
• 起止时间: 1999-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5251680?language=en
• 关键词: Funktionelle; Genomik; mit; mRNA; Protein

In recent years, technology for very fast automated genome sequencing has been developed at an amazing speed due to strong efforts in several genome projects. In contrast, development of methods for highly parallel investigation of the actual function of the genome`s translated message - the proteins - is in lagging far behind.This project will investigate a new strategy of functional interaction studies on proteins in a bio-combinatorial approach. Therefore, a method for the generation of mRNA-protein fusions, developed in Prof. Dr. Szostaks laboatory, will be combined with DNA microarray technology. Starting from a cDNA library, a corresponding mRNA-protein fusion library will be generated where every protein is covalently linked to its encoding mRNA. These fusions are subjected to an actvity selection procedure, specifically affinity to a certain protein target. As a result of the attached mRNA, the genetic information of the selectively enriched protein can be amplified by PCR an read out by hybridizing the fluorescently labeled PCR product with an appropriate DNA microarray. This strategy will allow screening for protein-protein and protein-small molecule interactions at genomic scale for virtually any organism. The method has great potential in the search for new drug targets an investigating metabolic pathways in general.

近年来，由于多个基因组项目的大力努力，极快速自动化基因组测序技术以惊人的速度发展。相比之下，对基因组翻译信息（蛋白质）的实际功能进行高度并行研究的方法的开发却远远落后。该项目将研究一种生物组合方法中蛋白质功能相互作用研究的新策略。因此，Szostaks 博士教授实验室开发的一种生成 mRNA-蛋白质融合体的方法将与 DNA 微阵列技术相结合。从 cDNA 文库开始，将生成相应的 mRNA-蛋白质融合文库，其中每个蛋白质都与其编码的 mRNA 共价连接。这些融合体经过活性选择程序，特别是对某种蛋白质靶标的亲和力。由于附着了 mRNA，选择性富集的蛋白质的遗传信息可以通过 PCR 进行扩增，并通过将荧光标记的 PCR 产物与适当的 DNA 微阵列杂交来读出。该策略将允许在几乎任何生物体的基因组规模上筛选蛋白质-蛋白质和蛋白质-小分子相互作用。该方法在寻找新药物靶点和研究一般代谢途径方面具有巨大潜力。