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The Tomato/GFP-FLP/FRT method for live imaging of mosaic adult Drosophila photoreceptor cells.

Author(s) : Dourlen P, Levet C, Mejat A, Gambis A, Mollereau B,
Journal : J Vis Exp
The Drosophila eye is widely used as a model for studies of development and neuronal degeneration. With the powerful mitotic recombination technique, elegant genetic screens based on clonal analysis have led to the identification of signaling pathways involved in eye development and photoreceptor (PR) differentiation at larval stages. We describe here the Tomato/GFP-FLP/FRT method, which can be used for rapid clonal analysis in the eye of living adult Drosophila. Fluorescent photoreceptor cells are imaged with the cornea neutralization technique, on retinas with mosaic clones generated by flipase-mediated recombination. This method has several major advantages over classical histological sectioning of the retina: it can be used for high-throughput screening and has proved an effective method for identifying thefactors regulating PR survival and function. It can be used for kinetic analysesof PR degeneration in the same living animal over several weeks, to demonstrate the requirement for specific genes for PR survival or function in the adult fly.This method is also useful for addressing cell autonomy issues in developmental mutants, such as those in which the establishment of planar cell polarity is affected.

Twist-DNA: computing base-pair and bubble opening probabilities in genomic superhelical DNA.

Author(s) : Jost D,
Journal : Bioinformatics
SUMMARY: Local opening of the DNA double helix is required in many fundamental biological processes and is, in part, controlled by the degree of superhelicity imposed in vivo by the protein machinery. In particular, positions of superhelically destabilized regions correlate with regulatory sites along the genome. Based on a self-consistent linearization of a thermodynamic model of superhelical DNA introduced by Benham, we have developed a program that predictsthe locations of these regions by efficiently computing base-pair and bubble opening probabilities in genomic DNA. The program allows visualization of results in standard genome browsers to compare DNA opening properties with other available datasets. AVAILABILITY AND IMPLEMENTATION: Source codes freely available for download at na, implemented in Fortran90 and supported on any Unix-based operating system (Linux, Mac OS X).