不同红色荧光蛋白在酵母细胞中的表达效果分析

Analysis of expression of the different red fluorescent proteins in yeast cells

目的比较不同红色荧光蛋白基因在酵母细胞中的表达效果。方法利用分子生物学方法,构建了4种红色荧光蛋白(DsRed)基因酵母报告载体,分别是p GPD-DsRed、p GPD-DsRed-express-2、p GPD-y DsRed和p GPDy DsRed-express-2,后两者含有酵母细胞偏好性的密码子,将4种DsRed酵母报告载体转入W303-1A酵母细胞,利用倒置荧光显微镜观察DsRed的表达,并用多功能酶标仪测酵母细胞的发光强度。结果各红色荧光发光强度明显不同,其中DsRed-express-2发光最强,其次是y DsRed-express-2,y DsRed发光强度最弱。结论在酵母细胞中红色荧光蛋白的强度与密码子偏好性无关;定量红色荧光蛋白表达强度最好选用DsRed-express-2报告基因。

Objective To compare the expression of fluorescent protein gene in yeast cells. Methods Four of red fluorescent protein（ DsRed） yeast expression vectors,pGPD-DsRed,pGPD-DsRed-express-2,pGPD-yDsred and pGPD-yDsred-express-2,were constructed using the molecular biology method. pGPD-yDsRed and pGPD-yDsRed-express-2contained the yeast cell preference codon. These 4 vectors were transformed into W303-1A yeast cells and DsRed expression was observed under the fluorescence microscope. Multifunctional microplate reader was used to measure the luminous intensity of yeast cells. Results The fluorescence intensities of the 4 the DsRed were significantly different.DsRed-express-2 emission was the strongest,followed by yDsRed-Express-2,and the weakest luminous intensity was yDsRed. Conclusion Red fluorescent protein strength has nothing to do with the codon preference in yeast cells. The best way is to select reporter gene of DsRed-express-2 to quantify the expression intensity of red fluorescent protein.