Modified bacterial artificial chromosomes for zebrafish transgenesis.

TitleModified bacterial artificial chromosomes for zebrafish transgenesis.
Publication TypeJournal Article
Year of Publication2006
AuthorsYang Z, Jiang H, Chaichanasakul T, Chachainasakul T, Gong S, Yang XWilliam, Heintz N, Lin S
JournalMethods
Volume39
Issue3
Pagination183-8
Date Published2006 Jul
ISSN1046-2023
KeywordsAnimals, Animals, Genetically Modified, Chromosomes, Artificial, Bacterial, Embryo, Nonmammalian, Escherichia coli, Gene Expression Regulation, Genes, Bacterial, Genes, Reporter, Genetic Engineering, Green Fluorescent Proteins, Luminescent Proteins, Plasmids, Rec A Recombinases, Recombinant Fusion Proteins, Recombination, Genetic, Zebrafish
Abstract

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.

DOI10.1016/j.ymeth.2006.04.011
Alternate JournalMethods
PubMed ID16828309
Grant ListRR013227 / RR / NCRR NIH HHS / United States