This proposal is designed to use the human cone transducin gene as a model for cone photoreceptor-specificgene expression. The components that are involved in the regulation mechanism are cis-acting elements and trans-acting factors. The cis-acting elements will be defined by transfecting retinoblastoma cells with an expression vector containing a reporter gene and a relevant DNA sequence. Trans-acting factors in the nuclei of retinoblastoma cells and in adult human retinas will be identified by DNase 1 footprinting and mobility shift assays. Protein-binding sites on cis-acting elements will be determined by methylation interference assays. The trans-acting factors identified will be cloned by screening a retina expression library with a specific cis-acting element. The identity of the trans-acting factors will be further confirmed by determining the partial amino acid sequences from the proteins or from the peptides. This work will generate new insights into gene expression in photoreceptors, and will provide a basis for the study of inherited retinal diseases at the molecular level.

Gene regulation of visual cycle and visual transduction proteins in human retina.

Special Interests
  • Vitamin A metabolism
  • Protein chemistry
  • Genetic engineering
  • Gene regulation
Genomics, proteomics
Biochemistry, Biochemistry, Proteins, Biological Sciences, Biology, Molecular, Eye Diseases, Gene Cloning, Gene Expression, Gene Regulation, Genetic Manipulation, Molecular Cloning, Molecular Probes, Nucleic Acid Sequencing, Sensory System, Tissue Culture, Transgenic Animals
Photoreceptor development
sensory system eye diseases blindness gene cloning genetic manipulation molecular cloning molecular probes nucleic acid sequencing gene regulation gene expression tissue culture genetically modified & transgenic animals ophthalmology biological sciences biochemistry & molecular biology vision biochemistry, proteins