Bacterial artificial chromosome transgenesis for zebrafish.
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.
Yang
Zhongan
Z
Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA, 90095-1606, USA.
Jiang
Hong
H
Lin
Shuo
S
D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000378:metabolism*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D030561:Databases, Nucleic Acid; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D000097573:Red Fluorescent Protein
D004247:DNA; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein