Reference - Detail
| RRC ID | 45912 |
|---|---|
| Author | Kage-Nakadai E, Imae R, Yoshina S, Mitani S. |
| Title | Methods for single/low-copy integration by ultraviolet and trimethylpsoralen treatment in Caenorhabditis elegans. |
| Journal | 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. |
| Volume | 68(3) |
| Pages | 397-402 |
| Published | 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 |
| Times Cited | 5 |
| WOS Category | BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Resource | |
| C.elegans | tm234 tm246 |