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Nucleic Acids Research, 1978, Vol. 5, No. 7 2643-2656
© 1978

Articles

Characterization of chick liver chromatin and analysis of its in vitro transcription products

Christa Dierks-Ventling1, Jürg Stalder2 and Johannes Gautschi3

1Friedrich Miescher-Institut P.O.Box 273, CH-4002 Basel, Switzerland 2Institut für allgemeine Mikrobiologie, Universität Bern Bern, Switzerland 3Institut für Pathologie, Universität Bern Bern, Switzerland

Received March 10, 1978. Carefully controlled preparation of chromatin from purified chick liver nuclei yielded over 50% native chromatin as shown by the analysis of the nucleosome pattern after micrococcal nuclease digestion. The size of DNA in this chromatin as analyzed on alkaline sucrose gradients varied from 10S to 19S, the majority being 14S. All endogenous RNA polymerases were represented in the chromatin preparation although to different extents: RNA polymerase I was the most and RNA polymerase II the least abundant. Initiation studies showed that endogenous RNA polymerase II was capable of initiating RNA chains during 5 min. Saturation of chromatin with purified homologous RNA polymerase II increased initiation to 10 min. The addition of heparin caused the RNA transcribed to be larger in size and of increased yield. Chromatin transcription with added purified RNA polymerase II in the presence of heparin produced RNA as large as 32S. A chromatin preparation of this kind would therefore be suitable to transcribe any eukary otic gene in vitro provided additional homologous RNA poly merase II is used.


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