一种高效干冻研磨破碎酵母细胞的方法

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、10min后破碎细胞的百分数为13.7%、30.1%、52.0%、85.1%、93.4%、94.6%、95.9%,0.1g酵母细胞研磨后抽提到的DNA和蛋白质的量分别为20μg和2.928mg。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。

A Simple and Efficient Method to Break Yeast Cell by Dry-frozen Grinding

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.