ZFIN ID: ZDB-PUB-090422-27
Bacterial artificial chromosome transgenesis for zebrafish
Yang, Z., Jiang, H., and Lin, S.
Date: 2009
Source: Methods in molecular biology (Clifton, N.J.)   546: 103-116 (Chapter)
Registered Authors: Jiang, Hong, Lin, Shuo, Yang, Zhongan
Keywords: Transgenic zebrafish, Bacterial artificial chromosomes, BAC modification, Fluorescent reporter genes, GFP/RFP
MeSH Terms:
  • Animals
  • Chromosomes, Artificial, Bacterial/metabolism*
  • DNA/genetics
  • DNA/metabolism
  • Databases, Nucleic Acid
  • Escherichia coli/genetics
  • Escherichia coli/metabolism
  • Gene Transfer Techniques*
  • Genes, Reporter
  • Genetic Engineering/methods
  • Green Fluorescent Proteins
  • Luminescent Proteins
  • Recombination, Genetic
  • Sequence Analysis, DNA
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
PubMed: 19378100 Full text @ Meth. Mol. Biol.
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.