MOLECULAR ANALYSIS OF TK AND HPRT GENE MUTATIONS IN L5178Y MOUSE LYMPHOMA CELLS

APPLIED GENETICS LABORATORIES, INC. PROPOSES TO UTILIZE NEWLY DEVELOPED MOLECULAR GENETIC TECHNIQUES TO ANALYZE THE SPECTRUM OF MUTAGENIC EVENTS THAT OCCUR AT THE TK AND HPRT LOCI IN MOUSE L5178Y CELLS. THE PURPOSE OF THESE STUDIES ISTO DETERMINE IF THE SPECTRUM OF GENETIC ABNORMALITIES REGISTERED AT THESE LOCI IS A FUNCTION OF THE HETEROZYGOUS/HOMOZYGOUS NATURE OF THE GENE LOCI INVOLVED, AND TO COMPARE THE SPECTRUM WITH THE KNOWN MUTAGENIC EVENTS THAT OCCUR IN HUMANS. BECAUSE SUCH MUTAGENIC EVENTS ARE IMPORTANT IN THE DEVELOPMENT OF HUMAN DISEASES (INCLUDING BIRTH DEFECTS AND CANCER), THE DEMONSTRATION OF A SIMILAR POTENTIAL FOR REGISTERING MUTAGENIC DAMAGE IN IN VITRO ASSAYS IS ESSENTIAL IN THE DESIGN OF MUTAGENESIS ASSAYS, ANDIS LIKELY TO CONTRIBUTE SIGNIFICANTLY TO THE UNDERSTANDING OF THE RELATIONSHIP BETWEEN IN VITRO MUTAGENIC POTENTIAL ANDHUMAN GENETIC DISEASE. PHASE I OF THIS PROJECT WILL DEAL WITH THE ADAPTATION OF RECENT MOLECULAR GENETIC TECHNICAL DEVELOPMENTS TO ANALYSIS OF MOUSE L5178Y CELLS (AT THE HPRT AND TK GENES), WHILE PHASE II WILL PROVIDE A FULL-SCALE ANALYSIS OF THE HEMIZYGOUS HPRT GENE AND THE TK GENE IN BOTH THE HETEROZYGOUS AND THE HEMIZYGOUS SITUATIONS.