Zebrafish Core

请参阅说明)： 斑马鱼核心(核心B)将为核心A和所有包含研究的项目提供直接支持 利用转基因斑马鱼。核心由迈克尔·帕森斯博士执导，他是一位经验丰富的调查员 斑马鱼基因，并将在第二年得到更多斑马鱼技术人员的支持。这个 具体目标如下：第一，提供集中胚胎显微注射服务 产生新的转基因品系；第二，为每一条转基因鱼提供专家日常护理 本提案中产生的服务，包括喂养和水箱维护、繁殖和基因分型服务； 第三，为成年斑马鱼和胚胎斑马鱼提供最先进的成像技术，包括共焦延时技术 显微镜。 核心B将负责所有野生型和遗传型的产生、繁育和维护 四个项目和核心A将使用的工程斑马鱼种群。这将包括每天 野生型胚胎的生产和Tol2转座子基元的胚胎显微注射 高效的转基因整合。在使用双顺反子转基因载体的帮助下，核心B也将 用于种系传递的筛选和用于产生独立品系的F1创始人。核心B 技术员还将负责日常喂食、水箱清洗、水质检测。核心B也将 协调成年鱼的养殖，以维护现有的鱼线和将鱼线杂交到 其他转基因背景。根据需要，从麻醉的鱼身上取下的鳍片将提供给 研究人员进行了PCR基因分型。最后，核心B将利用帕森斯博士广泛的 对活鱼进行共聚焦成像的经验，甚至可以描绘与之相关的微妙表型 具有胰腺编码和非编码候选癌基因的表达。 核心B被安置在一个新建成的1500平方米的房子里。英国《金融时报》共享斑马鱼设施。除了空间外， 胚胎注射和胚胎分选，该设施配备了34个机架的水生栖息地连续 具有连续水质监测和自动投药以保持恒定的流量系统 PH值和盐度。该系统的容量为-1400个2.5升水箱，可容纳30,000多名成年人 鱼。 相关性(请参阅说明)： 胰腺癌是人类最致命的恶性肿瘤之一，五年存活率为 低于5%，而且这一数字在过去40年中没有变化。通过确定人类的遗传基础 这种疾病，我们的计划将产生临床相关信息，很可能直接影响 化学预防、早期发现和治疗的战略。

Seeinstructions): The Zebrafish Core (Core B) will provide direct support to Core A and all projects incorporating studies utilizing transgenic zebrafish. The Core is directed by Dr. Michael Parsons, an experienced investigator in zebrafish genetics, and will be supported by additional zebrafish technicians by the second year. The following Specific Aims will be pursued: First, to provide centralized embryo microinjection services for generating novel transgenic lines; second, to provide expert daily care for each of the transgenic fish to be generated in this proposal, including feeding and tank maintenance, breeding and genotyping services; third, to provide state-of-the-art imaging of adult and embryonic zebrafish, including confocal time-lapse microscopy. Core B will be responsible for the generation, breeding and maintenance of all wild-type and genetically engineered zebrafish stocks to be utilized by the four projects and Core A. This will include daily production of wild-type embryos, and embryo microinjection of Tol2 transposon-based elements for high- efficiency transgene integration. Assisted by the use of bicistronic transgenic vectors, Core B will also screen for germline transmission and select F1 founders for generating independent lines. Core B technicians will also be responsible for daily feeding, tank cleaning, water quality testing. Core B will also co-ordinate breeding of adult fish, both for maintenance of existing lines and for crossing of lines onto other transgenic backgrounds. As needed, fin clips from anesthetized fish will be provided to investigators for PCR genotyping. Finally, Core B will take advantage of Dr. Parsons' extensive experience with confocal imaging of living fish, allowing delineation of even subtle phenotypes associated with pancreatic expression of coding and non-coding candidate cancer genes. Core B is housed in a newly completed 1500 sq. ft. Shared Zebrafish Facility. In addition to space for embryo injection and embryo sorting, the facility is outfitted with a 34-rack Aquatic Habitats continuous flow system with continuous water quality monitoring and automated dosing for maintenance of constant pH and salinity. The system has a capacity for -1400 2.5 liter tanks, and can house over 30,000 adult fish. RELEVANCE (See instructions): Pancreatic cancer represents one of the most deadly human malignancies, with five year survival rates of less than 5% and no change in this figure over the past four decades. By determining the genetic basis for this disease, our program will generate clinically relevant information that is likely to directly impact on strategies for chemoprevention, early detection and treatment.