論文 - 詳細
| RRC ID | 30452 |
|---|---|
| 著者 | Horn C, Offen N, Nystedt S, Häcker U, Wimmer EA. |
| タイトル | piggyBac-based insertional mutagenesis and enhancer detection as a tool for functional insect genomics. |
| ジャーナル | Genetics |
| Abstract |
Transposon mutagenesis provides a fundamental tool for functional genomics. Here we present a non-species-specific, combined enhancer detection and binary expression system based on the transposable element piggyBac: For the different components of this insertional mutagenesis system, we used widely applicable transposons and distinguishable broad-range transformation markers, which should enable this system to be operational in nonmodel arthropods. In a pilot screen in Drosophila melanogaster, piggyBac mutator elements on the X chromosome were mobilized in males by a Hermes-based jumpstarter element providing piggyBac transposase activity under control of the alpha1-tubulin promoter. As primary reporters in the piggyBac mutator elements, we employed the heterologous transactivators GAL4delta or tTA. To identify larval and adult enhancer detectors, strains carrying UASp-EYFP or TRE-EYFP as secondary reporter elements were used. Tissue-specific enhancer activities were readily observed in the GAL4delta/UASp-based systems, but only rarely in the tTA/TRE system. Novel autosomal insertions were recovered with an average jumping rate of 80%. Of these novel insertions, 3.8% showed homozygous lethality, which was reversible by piggyBac excision. Insertions were found in both coding and noncoding regions of characterized genes and also in noncharacterized and non-P-targeted CG-number genes. This indicates that piggyBac will greatly facilitate the intended saturation mutagenesis in Drosophila. |
| 巻・号 | 163(2) |
| ページ | 647-61 |
| 公開日 | 2003-2-1 |
| DOI | 10.1093/genetics/163.2.647 |
| PMID | 12618403 |
| PMC | PMC1462455 |
| MeSH | Animals DNA Transposable Elements* Drosophila melanogaster / genetics* Enhancer Elements, Genetic* Genes, Reporter Genomics* Mutagenesis, Insertional Mutation |
| IF | 4.015 |
| 引用数 | 113 |
| WOS 分野 | GENETICS & HEREDITY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 18 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ショウジョウバエ | DGRC#115635 |