Research objectives and content A new gene was identified in the host laboratory and preliminary n PP2/tkf. During embryonic development PP2/tkf is required for he correct formation of the muscle pattern and for the determination of the photoreceptor identify. We will analyse the structure of this gene and the nature of its protein product and will determine its position in the pathway leading to muscle and eye development in the larvae and the adult, respectively. To accomplish this we will: (1) Complete the characterization of the molecular organisation of the PP2/tkf locus as well as identication of the products encoded by the gene. (2) Study the function of PP2/tkf during embryonic development in the muscle and the tracheal systems. (3) Analyze the mechanism of action of PP2/tkf, studying the regulation of the gene as well as the biochemichal functions of the protein. (4) Correlate the different phenotypes that appear in the loss of function mutants with the function of the protein product.
Training content (objective, benefit and expected impact)
The training contents the project offers are primarily in bacterial, fly and yeast genetic engineering. The molecular characteriza- tion of the gene will require the application standard molecular genetic techniques. The analysis of the pattern of expression of the mRNA and protein in wild type and mutant animals will imply a fair amount of embryological analyses. The creation of new mutants will require the use o different mutangenic techniques as treatment with EMS, mobilization of P-elements,etc. The creation and use of antibodies will lead to the application of cellular biology techniques. The two hybrid system will be used to find molecules interacting with the gene.
The expected benefit of the project is a better knowledge of the gene, in regard of its structure, function and the genetic position during development in Drosophila. A human transcript homolog to PP2/tkf has been found recently. This makes the human gene a clear candidate to be responsible of a human disorder. Our work will lead to the possibility to identity what kind of genetic disorder is related to this gene, after following a strategy recently developed. Our final goal is to get basic information on the mechanisms of cell differentiation that we could eventually extrapolate to other organisms, including man.