title: Tol2-mediated transgenesis, gene trapping, enhancer trapping, and Gal4-UAS system.
abstract: The Tol2 element is an active transposon that was found from the genome of the Japanese medaka fish. Since the Tol2 transposition system is active in all vertebrate cells tested so far, it has been applied to germ line transgenesis in various model animals including fish, frog, chicken, and mouse, and to gene transfer in culture cells. In zebrafish, the Tol2 system consists of the transposase mRNA and a Tol2 transposon-donor plasmid, and is introduced into fertilized eggs by microinjection. Thus genomic integrations of the Tol2 construct are generated in the germ lineage and transmitted to the offspring very efficiently. By using the Tol2 transposition system, we have developed important genetic methods, such as transgenesis, gene trapping, enhancer trapping, and the Gal4-UAS system in zebrafish and applied to many aspects of biological studies. In this chapter, we describe how these methods are performed.
authors.affiliation: SOKENDAI (The Graduate University for Advanced Studies), Mishima, Shizuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kawakami
authors.affiliation: SOKENDAI (The Graduate University for Advanced Studies), Mishima, Shizuoka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Asakawa
authors.affiliation: SOKENDAI (The Graduate University for Advanced Studies), Mishima, Shizuoka, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Muto
authors.affiliation: SOKENDAI (The Graduate University for Advanced Studies), Mishima, Shizuoka, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Wada
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004251:DNA Transposable Elements / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D018014:Gene Transfer Techniques; D005854:Germ Cells / Q000254:growth & development; D009990:Oryzias / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D015027:Zebrafish / Q000235:genetics* / Q000254:growth & development
chemical_list: D004251:DNA Transposable Elements
keywords: Enhancer trapping; Gene trapping; The Gal4-UAS system; The Gal4FF trans-activator; The Tol2 transposable element; Transgenesis