Research objectives and content
The main goal of the present project is the elucidation of the molecular mechanisms involved in the establishment of the proximo-distal (PD) axis in the appendages of Drosophila. As a first step, we will study the expression pattern and genetic interactions of genes known to have a role in this process, and a preliminary hypothesis on the relationships of such genes will be suggested. As mutants for these genes do not affect all parts of the leg, some other genes must be necessarily involved. Thus, a second objective is the identification of new genes required for PD patterning among enhancer trap lines, according to their expression pattern from embryo to pupa, and the integration of their function in the framework proposed for the previously known genes. Alleles of these new genes will be isolated from the enhancer trap lines or by mutagenesis, in order to find mutant phenotypes of PD pattern. A molecular analysis will be performed for the most interesting gene found. Cloning will proceed from plasmid rescue of DNA adyacent to the enhancer trap insertion. The coding sequence will be identified in the candidate cytogenetical region by coincidence of the expression pattern of transcripts with the one of the enhancer traps. Then, the gene product will be sequenced to identify its nature.
Training content (objective, benefit and expected impact)
The work described in the project implies learning several new techniques that are necessary in the analysis of gene function and expression. The results and skills obtained will also be useful in further research projects in other organisms and genetic processes.