Core--Tansgenic fish facility

核心--转基因鱼设施

• 批准号: 6664592
• 负责人: Elwood Albert Linney
• 金额: $ 8.64万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-25 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6664592
• 关键词: Osteichthyes; animal colony; biomedical facility; genetically modified animals; reporter genes; zebrafish

This research core will facilitate the use of gene transfer and transgenes of fish in the different projects necessitating these technologies. The core will be setup to facilitate advice on transgenic construction, it will provide vectors and reporter genes, it will produce transgenic fish and screen the fish for the transgene. It will also provide instruction on handling and analyzing transgenic fish. Eventually, as new transgenic lines are produced that have firefly luciferase reporter genes, it will provide instructions on assaying the treated embryos and larvae for firefly induction by toxicants. In this context. the specific aims of this core will be: 1) To provide advice, vectors, cloned regulatory elements and reporter genes for the construction of transgenic vectors. For those laboratories not familiar with working with DNA, it will instruct individuals on how to make the transgenic constructs and/or make them for the laboratory. 2) To produce transgenic fish via DNA microinjection or pseudotyped retroviral vector infection. To screen the progeny of the potential transgenic founders to identify transgenic expressing progeny for the development of transgenic lines. To instruct individual, if desired, on the examination, breeding, and transgenic screening of the transgenic lines. 3) To collect and freeze down sperm from the transgenic lines for long- term storage. 4) To setup breedings so that embryos can be provided for treatment studies and to aid in the luciferase analysis of the IRES transgenic fish.

这一研究核心将促进在需要这些技术的不同项目中使用鱼类的基因转移和转基因。该核心将被设置为促进转基因构建的建议，它将提供载体和报告基因，它将产生转基因鱼和筛选转基因鱼。 它还将提供处理和分析转基因鱼的指导。最终，随着具有萤火虫荧光素酶报告基因的新的转基因品系的产生，它将提供关于测定经处理的胚胎和幼虫的毒物诱导萤火虫的指示。在这种情况下。该核心的具体目标是：1）为转基因载体的构建提供建议、载体、克隆调控元件和报告基因。对于那些不熟悉DNA工作的实验室，它将指导个人如何制作转基因构建体和/或为实验室制作它们。2)通过DNA显微注射或假型逆转录病毒载体感染生产转基因鱼。筛选潜在转基因建立者的后代，以鉴定转基因表达后代，用于转基因品系的开发。如果需要，指导个体进行转基因品系的检查、育种和转基因筛选。3)从转基因品系中收集精子并冷冻以供长期保存。4)建立胚胎库，以便为处理研究提供胚胎，并帮助IRES转基因鱼的荧光素酶分析。