Worm Breeder's Gazette 16(3): 13 (June 1, 2000)
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
CNRS-CGMC, University of Lyon, 69100 Villeurbanne, France
As an alternative to the systematic gene-directed knock-out program, we are exploring ways to create collections of transposon-tagged genes. Our strategy is to generate strains containing identified transposons insertions. Rubin and collaborators of the Drosophila gene disruption project are currently following a similar strategy with the fly genome (Genetics vol 153, p135). Over the last couple of years, we have run several pilot experiments making use of natural worm transposons to assay the feasibility of the strategy. In practice, we are following an approach which has been pioneered in the Plasterk lab. Starting from mut-7 or similar mutator backgrounds, we propagate the strain over a few generations to generate clones. Then, we determine the position of new insertions of Tc1, Tc3 and Tc5 by a modification of the transposon insertion display protocol described in the WBG (Vol 14, n!4 page 20), in which DNA flanking transposons can be amplified by anchored PCR, and sequenced. Over 500 clones have been generated in the course of the pilot experiments. In addition to insertions generated in our lab, a few clones have been contributed by Satis Sookhareea and Michel Labouesse. A total of 400 different insertions of Tc1, Tc3 and Tc5 have been identified (most clones we generate contain 0 to 2 insertions). The distribution of the insertions on the genetic map does not reveal any major bias. Over 50% of insertions located in annotated genome regions fall in genes. Introns tend to slightly favor insertions because of their high TA content, which is a target for the transposons we are following. The strains we produce are available to the C. elegans community upon request. Please consult our site at http://cgmc.univ-lyon1.fr/transposon%20project.htm for an updated list of lines and hits. Requests should be addressed to Laurent Segalat (email@example.com).