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（由申请人提供）：斑马鱼蛋白和抗体核心将迈出正式的第一步，创建国家资源，用于生产斑马鱼蛋白及其各自的抗体，供斑马鱼研究界使用。工作将集中在三项工作上：1）斑马鱼蛋白质和蛋白质片段的生产； 2）抗体片段的产生； 3) 阵列蛋白和亲和试剂的制造。斑马鱼的表达目标蛋白包括转录因子、酶、细胞骨架蛋白、细胞信号蛋白、生长因子、受体胞外域等。只要有可能，就会表达蛋白质的全长版本，但由于使用起来比较困难某些蛋白质类别（例如膜蛋白、多结构域蛋白），它们的表达将仅限于可溶性结构域。表达将主要在大肠杆菌中实现，昆虫细胞系统用于高优先级目标。第一年，我们将针对各种不同的蛋白质类别，以便我们可以开发与各种蛋白质相关的方案；我们的目标是表达 100 种蛋白质，并生成针对 50 个靶标的抗体。第二年，我们将靶点和抗体的生产规模扩大 50%，并逐步采用社区提名的蛋白质。预计第三年用户请求数量将增加一倍，总共 200 种蛋白质和 100 种抗体。此外，我们将制造蛋白质和抗体阵列，以便在第三年分发。为了监督这个试点项目，将设立一个由斑马鱼科学家组成的科学咨询委员会和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，包括一个包含可公开访问的数据的网站、科学会议上的演示以及所需试剂和方案的分发。