論文 - 詳細
| RRC ID | 42537 |
|---|---|
| 著者 | Kanayama N, Todo K, Takahashi S, Magari M, Ohmori H. |
| タイトル | Genetic manipulation of an exogenous non-immunoglobulin protein by gene conversion machinery in a chicken B cell line. |
| ジャーナル | Nucleic Acids Res |
| Abstract |
During culture, a chicken B cell line DT40 spontaneously mutates immunoglobulin (Ig) genes by gene conversion, which involves activation-induced cytidine deaminase (AID)-dependent homologous recombination of the variable (V) region gene with upstream pseudo-V genes. To explore whether this mutation mechanism can target exogenous non-Ig genes, we generated DT40 lines that bears a gene conversion substrate comprising the green fluorescent protein (GFP) gene as a donor and the blue fluorescent protein (BFP) gene as an acceptor. A few percent of the initially BFP-expressing cells converted their fluorescence from blue to green after culture for 2-3 weeks when the substrate construct was integrated in the Ig light chain locus, but not in the ovalbumin locus. This was the result of AID-dependent and the GFP gene-templated gene conversion of the BFP gene, thereby leading to the introduction of various sizes of GFP-derived gene segment into the BFP gene. Thus, G/B construct may be used to visualize gene conversion events. After switching off AID expression in DT40 cells, the mutant clones were isolated stably and maintained with their mutations being fixed. Thus, the gene conversion machinery in DT40 cells will be a useful means to engineer non-Ig proteins by a type of DNA shuffling. |
| 巻・号 | 34(2) |
| ページ | e10 |
| 公開日 | 2006-1-18 |
| DOI | 10.1093/nar/gnj013 |
| PII | 34/2/e10 |
| PMID | 16421270 |
| PMC | PMC1342040 |
| MeSH | Animals B-Lymphocytes / cytology B-Lymphocytes / immunology* Cell Line Chickens / genetics* Chickens / immunology Cytidine Deaminase / metabolism DNA Shuffling Gene Conversion* Genes, Immunoglobulin Light Chain Green Fluorescent Proteins / genetics Luminescent Agents Luminescent Proteins / genetics |
| IF | 11.502 |
| 引用数 | 20 |
| 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 | 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 | |
| ヒト・動物細胞 | DT40(RCB1464) |