論文 - 詳細
RRC ID | 45912 |
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著者 | Kage-Nakadai E, Imae R, Yoshina S, Mitani S. |
タイトル | Methods for single/low-copy integration by ultraviolet and trimethylpsoralen treatment in Caenorhabditis elegans. |
ジャーナル | Methods |
Abstract |
Single/low-copy transgene integration is essential for avoiding overexpression, ectopic expression and gene silencing in the germline. Here, we present an overview of a method that uses ultraviolet and trimethylpsoralen (UV/TMP) to generate single/low-copy gene integrations in Caenorhabditis elegans. Single/low-copy transgenes from extrachromosomal arrays are integrated into the genome using positive selection based on temperature sensitivity with a vps-45 rescue fragment and negative selection based on benzimidazole sensitivity with a ben-1 rescue fragment. The copy number of the integrated transgenes is determined using quantitative PCR. Our UV/TMP integration method, which is based on familiar extrachromosomal transgenics, provides a simple approach for generating single/low-copy gene integrations. |
巻・号 | 68(3) |
ページ | 397-402 |
公開日 | 2014-8-1 |
DOI | 10.1016/j.ymeth.2014.02.036 |
PII | S1046-2023(14)00091-7 |
PMID | 24613935 |
MeSH | Animals Animals, Genetically Modified / genetics* Caenorhabditis elegans / genetics* Genome / drug effects Genome / radiation effects Transgenes* / drug effects Transgenes* / radiation effects Trioxsalen / pharmacology Ultraviolet Rays |
IF | 3.812 |
引用数 | 5 |
WOS 分野 | BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
リソース情報 | |
線虫 | tm234 tm246 |