title: Modified bacterial artificial chromosomes for zebrafish transgenesis.
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.
authors.affiliation: Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA 90095-1606, USA.
authors.forename: Zhongan
authors.initials: Z
authors.lastname: Yang
authors.forename: Hong
authors.initials: H
authors.lastname: Jiang
authors.forename: Thawinee
authors.initials: T
authors.lastname: Chaichanasakul
authors.forename: Thawinee
authors.initials: T
authors.lastname: Chachainasakul
authors.forename: Shiaoching
authors.initials: S
authors.lastname: Gong
authors.forename: Xiangdong William
authors.initials: XW
authors.lastname: Yang
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Heintz
authors.forename: Shuo
authors.initials: S
authors.lastname: Lin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D004625:Embryo, Nonmammalian; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis; D008164:Luminescent Proteins / Q000032:analysis; D010957:Plasmids / Q000235:genetics; D011938:Rec A Recombinases / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis; D011995:Recombination, Genetic; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011938:Rec A Recombinases; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein