Modified bacterial artificial chromosomes for zebrafish transgenesis
Modified bacterial artificial chromosomes for zebrafish transgenesis
复制标题
DOI：
10.1016/j.ymeth.2006.04.011
复制
发表时间：
2006-07-01
期刊：
影响因子：
4.8
通讯作者：
Lin, Shuo
中科院分区：
文献类型：
--
作者：
Yang, Zhongan;Jiang, Hong;Lin, Shuo
使用细菌人工染色体（BAC）的转基因在指导外源基因的期望表达方面提供了更大的保真度。该技术在具有荧光蛋白报告子的光学透明斑马鱼中的应用使得能够对活生物体中的基因表达调控进行无与伦比的视觉分析。在这里，我们描述了一个简化的程序，直接选择多个BAC克隆的基础上，公共序列数据库，然后快速修改与GFP或RFP的转基因分析在斑马鱼。详细介绍了BAC DNA制备、斑马鱼胚胎显微注射和转GFP/RFP基因斑马鱼筛选的实验过程。(c)2006年爱思唯尔公司All rights reserved.
Transgenesis using bacterial artificial chromosomes (BAC) offers greater fidelity in directing desirable expression of foreign genes. Application of this technology in the optically transparent zebrafish with fluorescent protein reporters enables unparalleled visual analysis of regulation of gene expression in a living organism. Here we describe a streamlined procedure of direct selecting multiple BAC clones based on public sequence databases followed by rapid modification with GFP or RFP for transgenic analysis in zebrafish. Experimental procedures for BAC DNA preparation, microinjection of zebrafish embryos and screening of transgenic zebrafish carrying GFP/RFP modified BAC clones are detailed. (c) 2006 Elsevier Inc. All rights reserved.