Cloning Target Site for the DNA Binding Protein PAX 6

Our goal is to identify genes that are regulated by the putative transcription factor encoded by th(aniridia) locus. The specific aim of Phase I is to use bacterially produced pas-6 protein to identihuman genomic DNA. Isolation of sequences bound by pax-6 will be achieved by making fusion proteinsmaltose binding protein, and purifying protein/DNA complexes on amylose columns. Genomic fragments trecognition sites for pax-6 will be screened for the presence of sequences expressed in the retina aAt the end of Phase I, clones representing tissue specific targets of pax-6 binding will be obtainedfacilitate cloning of novel genes that are regulated by pax-6. Since PAX-6 and other PAX genes are khuman development disorders, cloning of target genes will permit the study of transcriptional mechanmolecular pathology of these disorders. In addition to furthering knowledge of disease mechanisms, timportant implications for manipulating gene expression in the nervous system.