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Fungal nuclei measure about 2 to 3 ^m in diameter and in the past their small size and poor staining by the conventional cytological stains discouraged their studies. However, recently several DNA-specific fluorescent dyes have been used for routine observations (Figure 2.1). In the last few years, a Neurospora chromosomal protein gene, histone H1, was tagged with a gene from a jellyfish that encodes a green fluorescent protein and glows bright green under blue light excitation (488 nm), thus making visualization of fungal nuclei possible under a fluorescence microscope. This method allows nuclei to be viewed in unfixed hyphae and to study the long distance movement of nuclei in living hyphae by video microscopy. The ease of isolating mutants has stimulated studies on the identification of nuclear genes involved in hyphal growth and morphogenesis. The feasibility of fusing fungal cells containing two different nuclear types into heter-okaryons makes possible the study of the cooperation or competition among nuclei in multinuclear hypha. Electrophoresis allows the separation of the tiny fungal chromosomes and makes the determination of the chromosome numbers more reliable than was possible by squash preparations.
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