Yang Zhongan, Jiang Hong, Lin Shuo
Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA, 90095-1606, USA.
Methods Mol Biol. 2009;546:103-16. doi: 10.1007/978-1-60327-977-2_7.
Transgenesis using bacterial artificial chromosomes (BAC) allows greater fidelity in directing desirable expression of transgenes. Application of this technology in the optically transparent zebrafish with fluorescent protein reporters enables unparalleled visual analysis of gene expression in a living organism. We have developed a streamlined procedure of directly selecting multiple BAC clones based on public sequence databases followed by rapid modification with green fluorescent protein or red fluorescent protein for transgenic analysis in zebrafish. In this chapter, experimental procedures for BAC DNA preparation and generation of transgenic zebrafish lines by microinjection are described.
使用细菌人工染色体（BAC）进行转基因可在指导转基因的理想表达方面实现更高的保真度。将该技术应用于带有荧光蛋白报告基因的光学透明斑马鱼，能够对活生物体中的基因表达进行无与伦比的可视化分析。我们基于公共序列数据库开发了一种直接选择多个BAC克隆的简化程序，随后用绿色荧光蛋白或红色荧光蛋白进行快速修饰，用于斑马鱼的转基因分析。在本章中，描述了BAC DNA制备以及通过显微注射生成转基因斑马鱼品系的实验程序。
