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Nucleic Acids Research, 1978, Vol. 5, No. 11 3993-4006
© 1978

Articles

Transcriptional properties of nucleoli isolated from Tetrahymena

Elmar Gocke, Johan Chr.Leer, Ole F. Nielsen and Ole Westergaard

Department of Molecular Biology, University of Aarhus 8000 Aarhus C, Denmark

Received September 25, 1978. Nucleoli can be isolated from Tetrahymena in a yield of 30–60%. The isolated nucleoli contain rDNA (at least 90% pure) and have a protein to DNA ratio of 30:1. The endogenous RNA-polymerase activity of the r-chromatin has the following properties: (i) The in vitro transcript has a maximal size identical to the in vivo 35S rRNA precursor, demonstrating correct termination on the gene, (ii) 79% of the in vitro transcript is complementary to cDNA of 17S and 25S rRNA which is close to the theoretical maximum for the 35S rENA precursor, (iii) the elongation rate of the endogenous RNA-polymerase molecules is 9–12 nucleotides/sec, (iv) an average of 4–16 active ENA polymerases are associated with each rDNA molecule depending upon the preparation.


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