A simple and efficient method of inducing targeted deletions in the drosophila genome

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• 作者：O. I. Kravchuk ; V. S. Mikhailov ; M. Yu. Savitsky
• 关键词：Drosophila ; method of deletion mutagenesis ; double ; strand DNA break repair
• 刊名：Russian Journal of Genetics
• 出版年：2015
• 出版时间：November 2015
• 年：2015
• 卷：51
• 期：11
• 页码：1144-1148
• 全文大小：542 KB
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• 作者单位：O. I. Kravchuk (1)
  V. S. Mikhailov (1)
  M. Yu. Savitsky (1) (2)
  
  1. Koltsov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, 119334, Russia
  2. Department of Molecular Biology, Umea University, Umea, 90187, Sweden
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Animal Genetics and Genomics
  Microbial Genetics and Genomics
  Russian Library of Science
  
• 出版者：MAIK Nauka/Interperiodica distributed exclusively by Springer Science+Business Media LLC.
• ISSN：1608-3369

文摘

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.