論文 - 詳細
| RRC ID | 28930 |
|---|---|
| 著者 | Kuspa A, Loomis WF. |
| タイトル | Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA. |
| ジャーナル | Proc Natl Acad Sci U S A |
| Abstract |
Introduction of restriction enzyme along with linearized plasmid results in integration of plasmid DNA at genomic restriction sites in a high proportion of the resulting transformants. We have found that electroporating BamHI or EcoRI together with pyr5-6 plasmids cut with the same enzyme stimulates the efficiency of transformation in Dictyostelium discoideum more than 20-fold over the rate seen when plasmid DNA alone is introduced. Restriction enzyme-mediated integration generates insertions into genomic restriction sites in an apparently random manner, some of which cause mutations. About 1 in 400 of the Dictyostelium transformants displayed arrested or aberrant development. The integrated plasmid, along with flanking genomic DNA, was excised from some of these mutants, cloned in Escherichia coli, and used to transform other Dictyostelium cells. Homologous recombination within the flanking sequences resulted in the same phenotypes displayed by the original mutants, directly demonstrating that the affected genes were responsible for the specific morphological phenotypes. This method of insertional mutagenesis should be useful for tagging, and subsequent cloning, of many developmentally important genes that can be identified by their mutant phenotypes. |
| 巻・号 | 89(18) |
| ページ | 8803-7 |
| 公開日 | 1992-9-15 |
| DOI | 10.1073/pnas.89.18.8803 |
| PMID | 1326764 |
| PMC | PMC50009 |
| MeSH | Animals Cell Aggregation Chromosome Deletion DNA Mutational Analysis DNA Restriction Enzymes / metabolism* DNA, Fungal / genetics* Dictyostelium / cytology Dictyostelium / genetics* Dictyostelium / growth & development Genes, Fungal* Mutagenesis, Insertional / methods* Plasmids Recombination, Genetic Restriction Mapping Transfection |
| IF | 9.412 |
| 引用数 | 406 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| 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 | 37 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 3.0 |
| リソース情報 | |
| 細胞性粘菌 | S00155 |