绿色荧光蛋白基因在裂殖酵母中的表达

将绿色荧光蛋白基因编码区序列克隆到大肠杆菌－裂殖酵母穿梭质粒ｐＲＥＰ３的ＢａｍＨⅠ￣ＳｍａⅠ位点，使之位于一个受硫胺素抑制的启动子和终止子的控制之下，用该质粒转化裂殖酵母，转化菌落于阳光下呈绿色，在３９５ｎｍ紫外光下发出强烈绿色荧光，在荧光显微镜下，用蓝光激发，可见该基因的表达明显受硫胺素的抑制，在没有选择压力的条件下，质粒丢失现象很严重，在完全培养基上，只有２０％的细胞发光，在丰富培养基上，发光

Expression of Green Fluorescent Protein Gene in Fission Yeast

Green fluolescent protein gene was cloned into the BamH I - Sma I sites of a E. colt - fission yeast shuttle plasmid pREP3 under control of nmtl promoter that is thiamine repressible. The plasmid resulted was used to transform fission yeast cells and the colonies of transformant have green color under sun light and strong green fluorescence under 395 nm UV. Under fluolescent microscope, yeast cells of transformant grown on selection meditun without thiamine all have very strong green fluorescence when blue excitation light is used, but the fluorescence is much weaker when thiamine is present in the medium. Only 20% of the cells have green fluorescence after die transformant cell forms a colony on a plate with complete medium. Less than 0. 1 % of the cells have green fluolescence after the transformant cell forms a colony on YES medium.