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FLP/FRT Induction of Mitotic Recombination in Drosophila Germline   

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Original research article

A brief version of this protocol appeared in:
eLIFE
Apr 2014

Abstract

The FLP/FRT system is a site-directed recombination technology based on the targeting of a recombination enzyme (flipase - FLP) to specific DNA regions designated as flipase recognition target (FRT) sites. Initially identified in Saccharomyces cerevisiae, the yeast FLP-enzyme and its FRT recombination targets were successfully transferred into each major chromosome arm in Drosophila (Golic and Lindquist, 1989). This offers the ability to mediate mitotic recombination in vivo during development in a controlled manner [revised in Theodosiou and Xu (1998)]. The controlled induction of the mitotic recombination events is usually performed by expressing the FLP under the control of the heat-shock (hs) promoter. This allows the expression of high FLP levels at specific developmental time windows. Strains carrying these genetically marked FLP/FRT chromosomes have greatly enhanced our ability to study gene function in both germline and somatic Drosophila tissues. Here we describe two different protocols: One to induce and identify homozygous mutant clones in ovaries and the other to generate female germline mutants for the analysis of maternal effects on embryogenesis.

Keywords: Drosophila, FLP/FRT, Mitotic Recombination, Germline

Copyright Prudêncio and Guilgur. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).
How to cite:  Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
  1. Prudêncio, P. and Guilgur, L. G. (2015). FLP/FRT Induction of Mitotic Recombination in Drosophila Germline. Bio-protocol 5(9): e1458. DOI: 10.21769/BioProtoc.1458.
  2. Guilgur, L. G., Prudencio, P., Sobral, D., Liszekova, D., Rosa, A. and Martinho, R. G. (2014). Requirement for highly efficient pre-mRNA splicing during Drosophila early embryonic development. Elife 3: e02181.
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