A simple and efficient method to break yeast cell by dry-frozen grinding | 一种高效干冻研磨破碎酵母细胞的方法_

Chinese. ［目的］建立简便有效的破碎酵母细胞的方法。［方法］离心收集酵母细胞，将培养液洗涤干净后直接将酵母细胞和研钵预冷至_-20 ℃，然后在冰冻状态将酵母细胞置于研钵中研磨，在相差显微镜下观察酵母细胞的形态，计算细胞破碎效率，并提取破碎酵母的染色体DNA和蛋白质。［结果］镜下观察研磨后酵母完整细胞数明显减少，并有大量细胞碎片。计数得出研磨1、2、3、4、6、8、10 min后破碎细胞的百分数为13.7%、30.1%、52.0%、85.1%、93.4%、94.6%、95.9%，0.1 g酵母细胞研磨后抽提到的DNA和蛋白质的量分别为20 μg 和2.928 mg。常规条件干冻酵母细胞直接研磨获得良好的破碎效果，破碎效率达95.9%。［结论］该研究建立了一种简便、高效而又经济的裂解酵母细胞的方法。_

English. ［Objective］ The research aimed to establish a simple and efficient method to break yeast cell. ［Method］Yeast cells were pre-frozen at -20 ℃ after collection and washing through centrifugation. Then under the frozen condition, the yeast cells were ground thoroughly with a mortar and pestle pre-frozen at -20 ℃. Following that, the ground yeasts were counted under a phase_contrast microscope. The gemomic DNA and the proteins of the yeasts were extracted with standard procedures. ［Result］ Lots of cell debris was observed after grinding and the number of broken entire yeasts was counted as 13.7%, 30.1%, 52_0%, 85.1%, 94.6%, 95.9% after grinding of 1, 2, 3, 4, 6, 8,10 min, respectively. 20 μg DNA and 2.928 mg protein were extracted from 0.1 g the ground yeast. High lysis percentage of yeast cells, up to 95.9%, was achieved by conventional grinding under dry-frozen condition. ［Conclusion The study constructs a simple, efficient and practical method was established to break yeast cells by dry-frozen grinding.