Core--Tansgenic fish facility

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。将建立核心，以便于提供转基因构建的建议，它将提供载体和报告基因，它将生产转基因鱼并对转基因鱼进行筛选。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转基因品系的产生，它将为检测毒物诱导萤火虫的处理胚胎和幼虫提供指导。在这种情况下。这个核心的具体目标将是：1)为转基因载体的构建提供建议、载体、克隆的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造转基因构建物和/或为实验室制造它们。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。筛选潜在转基因创建者的后代，鉴定转基因表达后代，为转基因品系的建立奠定基础。如有需要，指导个人进行转基因品系的检验、育种和转基因筛选。3)收集并冷冻转基因品系的精子，以供长期保存。4)建立育种方法，以便为治疗研究提供胚胎，并帮助进行IRES转基因鱼的荧光素酶分析。

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.