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Isolation and Properties of Macronuclei from Tetrahymena thermophila 1
Author(s) -
Corr Reinhard,
Eichler Wolfgang
Publication year - 1985
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1985.tb03011.x
Subject(s) - macronucleus , lysis , differential centrifugation , rna , dna , tetrahymena , biology , centrifugation , microbiology and biotechnology , alkaline lysis , digitonin , chemistry , biochemistry , chromatography , enzyme , gene , dna vaccination , plasmid
A simple and efficient method is described for the isolation of macronuclei from Tetrahymena thermophila (7B). The steps involved are deciliation and removal of the mucocysts’ contents by dibucaine treatment, digitonin mediated lysis, differential centrifugations, and finally isopyenic sucrose density gradient centrifugation. Judging from the distribution of marker enzymes and electron microscopy, the macronuclei obtained were free of cytoplasmic and paniculate contamination and were highly active in endogenous RNA‐synthesis (1.5 pmol UTP incorporation/ng DNA min at 30°C). The ratio of protein: RNA: DNA was 2.0:0.33:1.0 (weight) and each macronucleus contained an average of 17 pg DNA. The average yield of isolation was 50%.