Zebrafish Protein & Antibody Core

描述（由申请人提供）：斑马鱼蛋白质和抗体核心将采取第一个正式步骤，为斑马鱼蛋白质及其各自的抗体的生产创建国家资源，供斑马鱼研究界使用。研究工作将集中在三个方面：1）生产斑马鱼蛋白质和蛋白质片段; 2）生产抗体片段; 3）制造阵列蛋白质和亲和试剂。靶向表达的斑马鱼蛋白质包括转录因子、酶、细胞骨架蛋白、细胞信号传导蛋白、生长因子、受体的胞外域等。只要可能，将表达蛋白质的全长形式，但由于难以与某些蛋白质类别（例如，膜蛋白、多结构域蛋白），它们的表达将限于可溶性结构域。主要在E.大肠杆菌，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，并产生针对50种靶标的抗体。在第二年，我们将把目标和抗体的生产规模提高50%，并逐步采用社区提名的蛋白质。预计在第三年期间，用户请求的数量将翻一番，总共需要200种蛋白质和100种抗体。此外，我们将在第三年制造蛋白质和抗体的阵列。为了监督这个试点项目，将有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，包括一个网站，其中包含公开访问的数据，在科学会议上的演示文稿，并要求的试剂和协议的分布。

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.