SUMO fusion for genome-wide protein analysis

描述（由申请人提供）：从基因组测序项目中发现的大量新蛋白质为学术界和工业界的科学家创造了巨大的机会。为了了解新发现的蛋白质的结构，大规模的联盟和倡议正在进行中。不幸的是，蛋白质的表达和纯化是主要的瓶颈。细菌通常是重组蛋白表达的首选系统。在第一阶段，LifeSensors提出了一种新的sumo融合系统的发展，以提高大肠杆菌中多种蛋白质的表达和纯化。SUMO蛋白作为伴侣，促进包涵体蛋白的表达和溶解性增强。SUMO融合也被用作快速纯化伴侣蛋白的手柄。在第一阶段，进行了一项比较研究，以确定SUMO表达系统与其他领先融合技术的有效性。数据显示，SUMO系统在提高大肠杆菌蛋白的表达和溶解度方面优于5种主要融合系统。第一阶段的建议集中在酵母SUMO标签和SUMO蛋白酶上。在第二阶段，LifeSensors计划在其成功的基础上，通过识别新的酶和严格测试难以表达的蛋白质来扩大SUMO系统的范围。由于昆虫细胞系统已经发展成为真核表达的模型，因此在II期提出开发和销售昆虫细胞的SUMO系统。开发一种系统，增强蛋白质的表达，并提供基因组水平的快速纯化工具，将极大地帮助后基因组表达和蛋白质阵列技术。

DESCRIPTION (provided by applicant): A vast number of novel proteins discovered from genome sequencing projects are creating enormous opportunities for scientists in academia and industry. Large-scale consortia and initiatives are underway to understand the structures of newly discovered proteins. Unfortunately, protein expression and purification are a major bottleneck. Bacteria are usually the first system of choice for recombinant protein expression. In phase I, LifeSensors proposed the development of a novel SUMO-fusion system to enhance expression and purification of wide variety of proteins in E. coli. The SUMO protein acts as a chaperone to promote enhanced expression as well as solubility of inclusion body proteins. SUMO fusion is also used as a handle for rapid purification of partner proteins. In phase I, a comparative study was conducted to determine the effectiveness of the SUMO expression system with other leading fusion technologies. Data showed that the SUMO system was superior to the five leading fusion systems in enhancing expression and solubility of proteins in E. coli. The phase I proposal was centered on the yeast SUMO tag and SUMO protease. In phase II, LifeSensors proposes to build on its success to broaden the scope of SUMO system by identifying new enzymes and rigorously testing the system with difficuIt-to-express proteins. Since the insect cell system has developed as a model for eukaryotic expression, it is proposed in Phase II to develop and market a SUMO system for insect cells. Development of a system that enhances expression of proteins and offers rapid purification tools at the genomic level will greatly aid post-genomic expression and protein array technologies.