Preparation of Extracts from Yeast

INTRODUCTION Because yeast is exceptionally well suited to genetic analysis, both classical and molecular, it is an attractive system for expressing recombinant animal proteins for purification purposes. Methods available for lysing yeast cells include autolysis, pressure cells (e.g., French press), abrasives (glass bead vortexing), and enzymatic lysis (e.g., zymolase). One of the simplest methods, discussed in this protocol, involves the abrasive action of well-agitated glass beads. This is a very effective method for both low volumes (e.g., <1 mL using a microcentrifuge tube) and many liters using a specialized DynoMill apparatus. Cell breakage is typically >95%, as assessed by phase-contrast microscopy.