Zebrafish Core

参见说明）： 斑马鱼核心（核心B）将为核心A和所有纳入研究的项目提供直接支持 利用转基因斑马鱼。核心是由迈克尔帕森斯博士，一个经验丰富的调查员， 斑马鱼遗传学，并将在第二年得到更多斑马鱼技术人员的支持。的 以下具体目标将被追求：第一，提供集中的胚胎显微注射服务， 产生新的转基因品系;第二，为每一条转基因鱼提供专业的日常护理， 本提案中产生的，包括饲养和水箱维护、育种和基因分型服务; 第三，提供最先进的成年和胚胎斑马鱼成像，包括共聚焦时间推移 显微镜 核心B将负责所有野生型和遗传型的产生、育种和维持。 四个项目和核心A将利用的工程斑马鱼种群。这将包括每日 野生型胚胎的产生，以及基于Tol 2转座子的元件的胚胎显微注射， 高效转基因整合。通过使用双顺反子转基因载体的辅助，核心B还将 筛选生殖系传递并选择F1建立者以产生独立系。核心B 技术人员还将负责日常喂养，水箱清洗，水质检测。核心B还将 协调成鱼的繁殖，既为了维护现有的线路，也为了将线路穿越到 其他转基因背景。根据需要，将提供麻醉鱼的鳍夹， 研究人员进行PCR基因分型。最后，核心B将利用帕森斯博士广泛的 经验与共聚焦成像的活鱼，允许描绘甚至微妙的表型相关 与胰腺表达的编码和非编码候选癌基因。 核心区B位于一个新落成的1500平方米。英尺共享斑马鱼设施。除了空间， 胚胎注射和胚胎分选，该设施配备了一个34机架的水产养殖连续 连续水质监测和自动投加的流量系统， pH和盐度。该系统可容纳约1400个2.5升的水箱，可容纳30，000多名成年人 鱼. 相关性（参见说明）： 胰腺癌是最致命的人类恶性肿瘤之一，五年生存率为 不到5%，而且在过去40年中，这一数字没有变化。通过确定遗传基础， 这种疾病，我们的计划将产生临床相关的信息，可能会直接影响到 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.