Chapter 6 The Isolation of Nuclei from Fungi

Abstract

Publisher Summary This chapter describes the techniques for the isolation of nuclei from fungi. The chapter concentrates on the fungi, the true slime mold Physarum polycephalum , and the bread mold Neurospora crassa . The problem of isolating fungal nuclei and chromatin is that, the current methods might isolate proteins that appear to be histones when they are not. Quantitation of nuclear proteins that are not chromatin-bound nor as tightly bound as histones would be more difficult to establish. The use of snail gut enzyme to obtain protoplasts as intermediates would be an approach to overcome the problem of breaking the Neurospora cell wall. Some of the methods can be improved by use of inhibitors of proteases and nucleases and by alteration of metal ion concentrations. The combined use of protease and nuclease inhibitors, however, might produce artifacts because in mouse liver, such combinations resulted in anomalous changes in the fine structure of the nuclei.