Development of a new Chromosome-Identification Method and Gene-Mapping of Xenopus Laevis Chromosome.

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In order to identify Xenopus chromosomes and to make a gene-map of Xenopus chromosome, we tried to develop some methods, and cloning many chromosome specific genes, hormone genes, sex specific genes (or sex determining genes), and metamorphosis (or programmed cell death) marker genes.We developed ; 1. a highly sensitive nucleic acid detection method and gene-mapping method using chain reaction of fluorescence amplifiers, 2. a rapid isolation method for highly purified plasmid DNA, 3. a high resolution HPLC system using strong basic anion exchange resin (QA column), and 4. a DNA-immobilized anchor (DNA is enzymatically solidified by DNA ligase). By combination of these methods, we established ; 5. a new solid-phase cloning system, and 6. a hybrid selection method. According to use these method, we cloned ; 7. female specific DNAs (DNA markers of sex chromosome of Xenopus laevis, and 8. collagenase gene (a marker gene of metamorphosis in the amphibia). And, we are further cloning many chromosome specific gene markers and other marker genes. In addition to this research, we found an epidermal factor which stimulates collagenase production by fibroblasts in a reconstituted skin model, and clarified the hormonogenetic donor sites of thyroid hormone in thyroglobulin.