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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 different red fluorescent protein genes in yeast cells. METHODS: Four red fluorescent protein (DsRed) yeast reporter plasmids were constructed by using molecular biology method. They were p GPD-DsRed, p GPD-DsRed-express-2, p GPD-y DsRed and p GPDy DsRed-express- 2, the latter two contain yeast cell-preferred codons, the four DsRed yeast reporter vectors were transformed into W303-1A yeast cells, the inverted fluorescence microscope was used to observe the expression of DsRed, and the multi-function microplate reader was used to measure the luminescence intensity of yeast cells . The results showed that the fluorescence intensity of each red fluorescence was obviously different, among which DsRed-express-2 had the strongest luminescence, followed by y DsRed-express-2, and y DsRed had the weakest luminescence intensity. Conclusion The intensity of the red fluorescent protein in yeast cells is not related to the codon bias. The DsRed-express-2 reporter gene is the best choice for quantitative red fluorescent protein expression.