Abstract
Nuclei have important roles in DNA replication and gene expression of eukaryotes. Nuclei have been isolated and investigated in both plants and animals. On the other hand, there have been no reports on isolation of cell nuclei from fungi. Here we established a method for isolating and purifying nuclei from the mycelia of Zygomycota, Basidiobolus. To improve the yield and purity of nuclei, we investigated maceration methods, composition of buffers, and conditions of centrifugation, and established an optimum procedure. Basidiobolus cultured for 6 days in a potato-dextrose medium was enzymatically macerated, and then was homogenized in the presence of Triton X-100 and filtrated through a 20μm-openinng stainless sieve. The crude cell homogenate was loaded on a layer of a buffer containing 50% glycerol and centrifuged at 1500xg for 10min. The pellet contained pure nuclei. Observation under SEM revealed that the nuclei of Basidiobolus were spherical and not uniform in size.
