绿色荧光蛋白基因在裂殖酵母中的表达  

Expression of Green Fluorescent Protein Gene in Fission Yeast

作　　者：周炜[1] 罗忠训[1] 刘琳[1] 王庆平[1] 

机构地区：[1]湖北大学生命科学学院,湖北武汉430062

出　　处：《湖北大学学报（自然科学版）》1999年第1期73-75,共3页Journal of Hubei University：Natural Science

基　　金：国家自然科学基金!38970462;湖北省重点学科基金

摘　　要：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BamHISmaI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．用该质位转化裂殖酵母，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，发光细胞不到千分之一．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.

关 键 词：裂殖酵母 绿色荧光蛋白 基因表达 蛋白质 

分 类 号：Q753[生物学—分子生物学]