RRC ID 45912
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