The isolation of nuclei from the filamentous fungus Aspergillus nidulans. 1976

M A Gealt, and G Sheir-Neiss, and N R Morris

A method has been developed for isolating nuclei from the filamentous fungus Aspergillus nidulans. In this procedure, the mycelia from 14 to 16 h spore-derived cultures of A. nidulans NRI, a stable diploid strain, were frozen with liquid nitrogen and homogenized in a Waring blender. After homogenization, the mycelia were warmed to 4 degrees C and the nuclei were purified from the homogenate by differential centrifugation followed by sedimentation through 2.1 M-sucrose. The final nuclear yield was 15 to 20%, based on DNA estimations. The purified nuclear pellet was free of whole cells. The morphology of the isolated nuclei resembled that of the in situ nuclei; they contained a nucleolus, chromatin, and had a surrounding double membrane. The purified nuclei were characterized by a DNA:RNA:protein ratio of 1:2.8:8.7.

UI MeSH Term Description Entries
D002458 Cell Fractionation Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS. Cell Fractionations,Fractionation, Cell,Fractionations, Cell
D002467 Cell Nucleus Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed) Cell Nuclei,Nuclei, Cell,Nucleus, Cell
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D005656 Fungal Proteins Proteins found in any species of fungus. Fungal Gene Products,Fungal Gene Proteins,Fungal Peptides,Gene Products, Fungal,Yeast Proteins,Gene Proteins, Fungal,Peptides, Fungal,Proteins, Fungal
D001233 Aspergillus nidulans A species of imperfect fungi from which the antibiotic nidulin is obtained. Its teleomorph is Emericella nidulans. Aspergillus nidulellus,Emericella nidulans
D012313 RNA A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed) RNA, Non-Polyadenylated,Ribonucleic Acid,Gene Products, RNA,Non-Polyadenylated RNA,Acid, Ribonucleic,Non Polyadenylated RNA,RNA Gene Products,RNA, Non Polyadenylated
D013347 Subcellular Fractions Components of a cell produced by various separation techniques which, though they disrupt the delicate anatomy of a cell, preserve the structure and physiology of its functioning constituents for biochemical and ultrastructural analysis. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p163) Fraction, Subcellular,Fractions, Subcellular,Subcellular Fraction

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