斑马鱼转基因平台的建立  

Foundation of Platform for Zebrafish Transgenic Technology

作　　者：刘丽丽[1] 王健[1] 王海胜[1] 余凯敏 李国超[1] 闫艳春[1] 

机构地区：[1]中国农业科学院研究生院,北京100081

出　　处：《生物技术通报》2013年第10期120-126,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31170119);中国农业科学院基础研究基金项目(0042011006;0042012003)

摘　　要：建立实验室斑马鱼转基因平台,旨在为实验室斑马鱼规模化繁殖、转基因斑马鱼研究提供技术流程、生物材料和执行标准。以一对Tübingen品系斑马鱼为材料,在实验室条件下进行扩大繁殖;将广谱表达绿色荧光蛋白GFP真核载体pEGFPN3通过显微注射的手段转入斑马鱼胚胎,获得转基因斑马鱼;通过荧光显微镜观察斑马鱼GFP表达情况,并初步筛选转基因个体;以观察到GFP表达的斑马鱼为材料,利用PCR验证外源基因pEGFP-N3在宿主细胞内的整合情况。结果显示,在实验室内建立了完善的斑马鱼养殖系统,可以长期进行斑马鱼饲养、繁殖、育种等研究工作;建立了斑马鱼基础分子生物学试验体系,可以进行斑马鱼分子水平的研究工作;建立了斑马鱼转基因平台,为构建转基因斑马鱼品系、外源基因整合机制等研究奠定基础。The aim was to provide biomaterial, technique flow and protocol standard of transgenic zebrafish study via founding a platform of zebrafish transgenic technology. A couple of Tfibingen zebrafish were muhiplicated whose zygotes were microinjected with high active constitute plasmid pEGFP-N3. Injected eggs were collected and cultured and further observation. Fluorescent microscopic observation was followed by PCR on these embryos to screen and certify GFP expressed transgenic zebrafish. Results showed that the present study developed an intact cultivation system of zebrafish including its cultivation, reproduction and breeding. We also dug out the common emergency and proposed possible solutions during zebrafish maintenance. We designed the molecular processes with zebrafish embryo as material in our lab. Especially a platform of zebrafish transgenic technology was founded for the regeneration of transgenic zebrafish lines and study of transgene integration mechanism.

关 键 词：斑马鱼 繁殖 转基因平台 显微注射 

分 类 号：X174[环境科学与工程—环境科学]