Secretion of Ubiquitin-fusion Proteins

基因组学项目正在发现大量的新基因，因此需要表达用于治疗、诊断和理解蛋白质结构和功能的新蛋白质。LifeSensors提出了一种新的泛素融合系统来解决这一需求。该方案利用新发现的泛素调控特性将重组蛋白分泌到培养基中。该融合系统将在杆状病毒和酵母蛋白表达和分泌系统中使用不同结构和功能的已知蛋白进行测试。我们预计，结合分泌物的ub融合技术的改进将显著可靠地提高在其他系统中难以表达和纯化的生物活性蛋白的生产和纯化。这些结果将在二期中用于开发表达多种蛋白质的通用试剂盒或作为专门cDNA文库表达系统的一部分。拟议的商业应用：由于缺乏一种在真核或原核系统中适用于结构多样化蛋白质的简单方法，用于研究和生物制药目的的生物活性蛋白的表达受到阻碍。我们建议使用泛素融合技术结合分泌来提高生物活性蛋白的定量和定性表达和后续纯化。结合这些改进的试剂盒的生产可能会导致一种更可靠的方法来表达和纯化所有生物体的蛋白质。

Genomics programs are discovering a large number of novel genes thus creating a need to express new proteins for therapeutics, diagnostics and to understand protein structure and function. LifeSensors proposes the creation of a new ubiquitin-fusion system to address this need. This proposal makes use of newly discovered regulatory properties of ubiquitin to secrete recombinant proteins into the medium. This fusion system will be tested in the baculovirus and yeast protein expression and secretion systems using known proteins of varied structure and function. It is expected that the improvements to the ub-fusion technology combined with secretion will significantly and reliably enhance the production and purification of biologically active proteins that have been difficult to express and purify in other systems. These results will be used in Phase II to develop a generic kit for expressing a variety of proteins or as part of specialized cDNA library expression systems. PROPOSED COMMERCIAL APPLICATIONS: Expression of biologically active proteins for research and biopharmaceutical purposes is hampered by the lack of a simple method that works for structurally diverse proteins in either eukaryotic or prokaryotic systems. We propose the use of ubiquitin-fusion technology combined with secretion to enhance the quantitative and qualitative expression and subsequent purification of biologically active proteins. The production of a kit incorporating these improvements could lead to a more reliable method of expressing and purifying proteins from all organisms.