Analysis of genes and genome by the tol2-mediated gene and enhancer trap methods.
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.
Division of Molecular and Developmental Biology, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka, 411-8540, Japan.
Akihiro
A
Urasaki
Koichi
K
Kawakami
D000818:Animals; D030801:Animals, Genetically Modified; D003433:Crosses, Genetic; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods*; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D016254:Mutagenesis, Insertional / Q000379:methods*; D011817:Rabbits; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D004247:DNA