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- Title
Characterization of an exchangeable gene trap using pU-17 carrying a stop codon-βgeocassette.
- Authors
Taniwaki, Takuya; Haruna, Kyoko; Nakamura, Hiroshi; Sekimoto, Tomohisa; Oike, Yuichi; Imaizumi, Takashi; Saito, Fumiyo; Muta, Mayumi; Soejima, Yumi; Utoh, Ayako; Nakagata, Naomi; Araki, Masatake; Yamamura, Ken-ichi; Araki, Kimi
- Abstract
We have developed a new exchangeable gene trap vector, pU-17, carrying the intron-lox71-splicing acceptor (SA)-βgeo-loxP-pA-lox2272-pSP73-lox511. The SA contains three stop codons in-frame with the ATG ofβgalactosidase/neomycin-resistance fusiongene (βgeo) that can function in promoter trapping. We found that the trap vector was highly selective for integrations in the introns adjacent to the exon containing the start codon. Furthermore, by using the Cre-mutantloxsystem, we successfully replaced theβgeogene with theenhanced green fluorescent protein(EGFP) gene, established mouse lines with the replaced clones, removed the selection marker gene by mating with Flp-deleter mice, and confirmed that the replacedEGFPgene was expressed in the same pattern as theβgeogene. Thus, using this pU-17 trap vector, we can initially carry out random mutagenesis, and then convert it to a gain-of-function mutation by replacing theβgeogene with any gene of interest to be expressed under the control of the trapped promoter through Cre-mediated recombination.
- Subjects
GENETIC mutation; MUTAGENESIS; INTRONS; NEOMYCIN; GREEN fluorescent protein; AMINOGLYCOSIDES
- Publication
Development, Growth & Differentiation, 2005, Vol 47, Issue 3, p163
- ISSN
0012-1592
- Publication type
Article
- DOI
10.1111/j.1440-169X.2005.00792.x