NAME: Adam Short
PROJECT TITLE: Genomic Comparison of Acanthmaoeba Isolates from Diverse Environments.

Acanthamoeba spp. are free-living amoebae found in aquatic and soil environments worldwide. Some strains are known to be pathogenic causing the sight-threatening infection, amoebic keratitis, or granulomatous amoebic encephalitis, a frequently fatal, opportunistic infection in immunocompromised individuals such as AIDS patients. Many strains of Acanthamoeba have been isolated and characterized from a variety of environments and have been shown to differ widely in their tolerance to various temperatures, salinities, and pH. Exactly what accounts for these observed differences as well as whether or not they are pathogenic, is unknown. In this project, Acanthamoeba isolates will be selected for whole genome analysis based on their physiological characteristics and pathogenic potential. Intact chromosomes will be separated by CHEF gel electrophoresis to generate electrophoretic karyotypes.  Whole chromosome separations will then be restriction digested in situ and subjected to 2nd dimension agarose gel electrophoresis to generate ‘genomic fingerprints’ which will be compared among the different isolates. Any unique bands may represent genes that contribute to an isolate’s ability to survive under specific environmental conditions or contribute to their ability to cause infection.