Development of systems for cell culture and expression vector to generate transgenic marine invertegra

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1. Identification of cis-active elements that regulate expression of sea urchin gene :Several sequence elements that regulate transcription of the sea urchin Ars gene were detected by reporter assay around its transcription start site. Among them between Sox-binding sitebetween -168bp and -162bp is most prominent element. Around -186bp cis-element that represses the promoter activity was detected.2. Validity of insulator element in gene transfer experiments :An insulator element present at the upstream of the sea urchin Ars gene showed insulator activity also in cells of Drosophila, mouse and humane3. Cell culture system for mantle epitermal cells of pearl oyster :Primary cell culture system for mantle epitermal cells of pearl oyster was established, though they do not proliferate in vitro.4. Culturing of sea urchin embryonic cells :Though we succeeded in establishing in vitro culture of cells from sea urchin pluteus larvae, we failed to culture them beyond three days because of difficulty in preventing proliferation of contaminating fungi.