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A simple and efficient method of inducing targeted deletions in the drosophila genome
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
2015 (English)In: Russian journal of genetics, ISSN 1022-7954, E-ISSN 1608-3369, Vol. 51, no 11, 1144-1148 p.Article in journal (Refereed) Published
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Abstract [en]

Deletion mutagenesis is one of the most efficient approaches to studying gene function. However, conventional methods of inducing targeted mutations in the drosophila genome are timeand labor-consuming. This work proposes a new, simple, and effective method of producing drosophila mutants with gene deletions. The method involves the insertion of I-SceI and I-CreI recognition sites and a fragment homologous to the target sequence into the chromosome region of interest by means of an attB-containing construct, the induction of double-strand DNA breaks by the appropriate meganuclease, and their repair by homologous recombination. The procedure results in a deletion extending from the attP-site to the target locus. A cassette was designed to enable single-step construct production for the deletion of any given genomic region. A set of markers facilitates the selection of recombination events. The efficacy of the proposed technique was confirmed by the induction of a 47-kb deletion containing the qtc gene.

Place, publisher, year, edition, pages
2015. Vol. 51, no 11, 1144-1148 p.
Keyword [en]
Drosophila, method of deletion mutagenesis, double-strand DNA break repair
National Category
Genetics
Identifiers
URN: urn:nbn:se:umu:diva-113442DOI: 10.1134/S1022795415110101ISI: 000365129000012OAI: oai:DiVA.org:umu-113442DiVA: diva2:885178
Available from: 2015-12-18 Created: 2015-12-18 Last updated: 2015-12-18Bibliographically approved

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Savitsky, Mikhail
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Department of Molecular Biology (Faculty of Medicine)
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CiteExportLink to record
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