Separation and purification of macronuclei from macronuclear fragments and micronuclei in the ciliate Paramecium aurelia

• Published in: Experimental cell research Vol. 90; no. 1; pp. 143 - 152
• Main Authors: Skoczylas, B., Soldo, A.T.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 1975
• Subjects: Acid Phosphatase - metabolism; Amino Acids - metabolism; Animals; Carbon Radioisotopes; Cell Division; Cell Fractionation; Cell Nucleus - metabolism; Cell Nucleus - ultrastructure; Centrifugation, Density Gradient; Deoxyribonucleases - metabolism; DNA - analysis; DNA Nucleotidyltransferases - metabolism; DNA-Directed RNA Polymerases - metabolism; Glucosephosphate Dehydrogenase - metabolism; Microscopy, Electron; Microscopy, Phase-Contrast; Nucleoproteins - analysis; Paramecium - growth & development; Paramecium - metabolism; Paramecium - ultrastructure; Resins, Plant; Ribonucleases - metabolism; RNA - analysis; Sucrose
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/0014-4827(75)90367-5

A method has been developed for the isolation and purification of macronuclei from the ciliated protozoan Paramecium aurelia, free from contamination by macronuclear fragments, micronuclei and cellular debris. It is based on differential centrifugation in discontinuous gradients prepared from sucrose-gum arabic solutions. Macronuclei prepared by this procedure possessed both DNA polymerase and RNA polymerase activity and were capable of incorporating amino acids into an acid-insoluble fraction in vitro. They contained an average of 90 pg of DNA, 94 pg of RNA and 189 pg of protein per nucleus.