Zebrafish Protein & Antibody Core

斑马鱼蛋白

• 批准号: 7125256
• 负责人: BRIAN KENNETH KAY
• 金额: $ 28.94万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7125256
• 关键词: Escherichia coli; affinity labeling; animal genetic material tag; antibody; biotechnology; chemical registry /resource; gene expression; immunologic substance development /preparation; microarray technology; oligopeptides; peptide chemical synthesis; peptide library; reagent /indicator; zebrafish

DESCRIPTION (provided by applicant): The Zebrafish Protein & Antibody Core will take the first formal step in creating a national resource for the production of zebrafish proteins and their respective antibodies for use by the zebrafish research community. Effort will center on three efforts: 1) production of zebrafish proteins and protein fragments; 2) production of antibody fragments; and 3) the fabrication of arrayed proteins and affinity reagents. Zebrafish proteins targeted for expression would include transcription factors, enzymes, cytoskeletal proteins, cell signaling proteins, growth factors, ectodomains of receptors, etc. Wherever possible, full-length versions of the proteins would be expressed, but due to the difficulty of working with certain protein classes (e.g., membrane proteins, multi-domain proteins), their expression would be limited to soluble domains. Expression would be achieved primarily in E. coli, with insect cell systems used for high priority targets. For the first year, a variety of different protein classes would be targeted so that we may develop protocols relevant to a wide range of proteins; we would aim for the expression of 100 proteins, and the generation of antibodies to 50 targets. During the second year, we would ramp up the production scale of targets and antibodies by 50%, and phase in proteins nominated by the community at large. It is anticipated that the number of user requests would double during the third year, for a total of 200 proteins and 100 antibodies. In addition, we would fabricate arrays of proteins and antibodies for distribution during the third year. To oversee this pilot project, there will be a scientific advisory committee composed of zebrafish scientists, and a technical advisory committee composed of biotechnologists. We propose a sharing plan consisting of a website containing publicly accessible data, presentations at scientific meetings, and distribution of requested reagents and protocols.

描述(由申请者提供)：斑马鱼蛋白和抗体核心将迈出正式的第一步，创建一个国家资源，生产斑马鱼蛋白及其相应的抗体，供斑马鱼研究社区使用。这项工作将集中在三个方面：1)斑马鱼蛋白质和蛋白质片段的生产；2)抗体片段的生产；3)阵列蛋白质和亲和试剂的制造。斑马鱼的目标蛋白将包括转录因子、酶、细胞骨架蛋白、细胞信号蛋白、生长因子、受体的胞外结构域等。在可能的情况下，将表达全长版本的蛋白质，但由于难以与某些蛋白质类别(如膜蛋白、多结构域蛋白)一起工作，它们的表达将限于可溶性结构域。表达将主要在大肠杆菌中实现，昆虫细胞系统将用于高优先级靶标。在第一年，各种不同的蛋白质类别将成为目标，以便我们可以制定与广泛的蛋白质相关的方案；我们的目标是表达100种蛋白质，并产生针对50个目标的抗体。在第二年，我们将把靶标和抗体的生产规模扩大50%，并逐步引入社会各界提名的蛋白质。预计在第三年，用户请求的数量将翻一番，总共有200个蛋白质和100个抗体。此外，我们将制作蛋白质和抗体阵列，以便在第三年分发。为了监督这一试点项目，将成立一个由斑马鱼科学家组成的科学咨询委员会，以及一个由生物技术专家组成的技术咨询委员会。我们提出了一个共享计划，其中包括一个网站，其中包含公开可访问的数据、在科学会议上的陈述以及所请求的试剂和方案的分发。