Investigation of Unutilized Bioluminescent Systems and Fluorescent Proteins, and Development of New Molecular Proves.

1)从海洋发光蠕虫中，Odontosyll是从美国圣地亚哥Mission Bay收集的磷光物质，在荧光素-荧光素酶反应的指导下，分离出发光物质Odontosyll是荧光素。分离得到的荧光素和粗荧光素酶的生物发光光谱（λmax 500nm）与活体荧光素相似。紫外线光谱基本上与Shimomura先前报告的相同。高分辨ESI质谱测定荧光素分子式为C_<15>H_8O_<12>S_3.2)在新喀里多尼亚对海洋发光生物进行了探索，发现了一些发光蠕虫和海兔。3)采用荧光素-荧光素酶反应的方法，从新西兰皮隆尼亚山附近的努图努溪中采集的淡水帽贝如蜗牛Latia neritoides中分离出荧光素酶。详细的生化研究表明，荧光素酶的活性形式是一个分子质量为31kDa的n链糖蛋白单体六聚体。荧光素酶基因克隆成功。但该基因的异源表达尚未成功。

1)From the marine luminescent worm, Odontosyll is phosphorea collected off from Mission Bay, San Diego, U.S.A., the light producing substance, Odontosyll is luciferin was isolated under the guidance of luciferin-luciferase reaction. The bioluminescence spectrum (λmax 500nm) using the isolated luciferin and the crude luciferase was similar to that of live specimens. The UV spectrum was essentially identical with that reported previously by Shimomura. The molecular formula of the luciferin was determined from the high resolution ESI mass spectrum to be C_<15>H_8O_<12>S_3.2)Exploration of luminescent marine organisms was performed at New Caledonia, some luminous worm and sea hare was found.3)From the fresh-water limpet like snail, Latia neritoides collected at the Nugtunu stream near Mt. Pirongia, New Zealand, the luciferase was isolated under the guidance of luciferin-luciferase reaction. The detailed biochemical studies indicated that the active form of the luciferase was found to be a hexamer of N-linked glycoprotein monomer having the molecular mass 31kDa. The cloning of the luciferase gene was succeeded. However heterologous expression of the gene was not succeeded so far.