A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode Caenorhabditis elegans

The generation of a large collection of defined transposon insertion mutants is of general interest to the Caenorhabditis elegans research community and has been supported by the European Union. We describe here a semi-automated high-throughput method for mutant production and screening, using the heterologous transposon Mos1. The procedure allows routine culture of several thousand independent nematode strains in parallel for multiple generations before stereotyped molecular analyses. Using this method, we have already generated >17 500 individual strains carrying Mos1 insertions. It could be easily adapted to forward and reverse genetic screens and may influence researchers faced with making a choice of model organism

The COPAS Profiler can discriminate between worms carrying different transgenic arrays and can be used to sort worms of a desired genotype

<p><b>Copyright information:</b></p><p>Taken from "A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode "</p><p></p><p>Nucleic Acids Research 2006;35(2):e11-e11.</p><p>Published online 12 Dec 2006</p><p>PMCID:PMC1903375.</p><p>© 2006 The Author(s).</p> Fluorescent micrographs (upper panels) and profiles (lower panels) for individual worms carrying the substrate array (), the transposase array (), associated with specific GFP expression in the pharynx and in the coelomocytes, respectively, or both (). Fluorescence is measured in arbitrary but constant units. Time-of-flight is a measure of length ()

Successive steps in image analysis to determine whether a well contains worms

<p><b>Copyright information:</b></p><p>Taken from "A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode "</p><p></p><p>Nucleic Acids Research 2006;35(2):e11-e11.</p><p>Published online 12 Dec 2006</p><p>PMCID:PMC1903375.</p><p>© 2006 The Author(s).</p> An image of each entire well of a 96-well plate was acquired automatically () and processed in successive steps using the ImageJ (NIH) software: overall light and contrast enhancement (), loading of a circular region and inversion of gray level at each pixel inside the region (), binarization on the basis of gray level at each pixel in the entire image (), removal of all the selected particles outside the loaded region () and removal of all the particles shorter than a minimum size ()

Utilisation de PROLOG, langage d'intelligence artificielle, en biologie moleculaire

CNRS AR 11930 / INIST-CNRS - Institut de l'Information Scientifique et TechniqueSIGLEFRFranc