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The Tol2 transposon system can create insertions in the zebrafish genome efficiently. By using this system, the gene trap and enhancer
trap methods have been developed. The gene trap and enhancer trap constructs contain the green fluorescent protein (GFP) reporter
gene or the yeast Gal4 transcription activator gene. By creating random integrations of these constructs in the genome, transgenic
fish expressing the GFP gene or the Gal4 gene in specific cells, tissues or organs are generated. These fish are valuable
resources for developmental biology. Especially, the Gal4-expressing transgenic fish can be used to ectopically express any
gene of interest placed downstream of the Gal4 recognition sequence, UAS, and thereby allow visualization, modification or
ablation of the Gal4-expressing cells. In this chapter, we will describe how the gene trap and enhancer trap screens can be
performed and how the transposon insertions created by these methods can be analyzed.
Affiliation(s): (4) Division of Molecular and Developmental Biology, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka 411-8540, Japan
Book Title: Zebrafish: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 546 | Pub. Date: Jan-01-2010 | Page Range: 85-102 | DOI: 10.1007/978-1-60327-977-2_6
Subject: Biochemistry
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