Funktionelle Genomik mit mRNA-Protein-Konjugaten und DNA Mikrochips

近年来，由于几个基因组计划的大力努力，非常快速的自动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平行研究的方法的发展远远落后。本项目将探索一种生物组合方法研究蛋白质功能相互作用的新策略。因此，Szostaks教授实验室开发的一种生成mrna -蛋白融合体的方法将与DNA微阵列技术相结合。从cDNA文库开始，生成相应的mRNA-蛋白融合文库，其中每个蛋白与其编码mRNA共价连接。这些融合受到活性选择程序的影响，特别是对特定蛋白质目标的亲和力。由于附着的mRNA，选择性富集蛋白的遗传信息可以通过PCR扩增，并通过将荧光标记的PCR产物与合适的DNA微阵列杂交读出。这一策略将允许在几乎任何生物体的基因组尺度上筛选蛋白质-蛋白质和蛋白质-小分子相互作用。该方法在寻找新的药物靶点和研究代谢途径方面具有很大的潜力。

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.