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Nucleic Acids Research, 1980, Vol. 8, No. 3 573-586
© 1980

Articles

Extraction of transcribing SV4O chromosomes in very low salt

Howard J. Edenberg

Department of Biochemistry, Indiana University School of Medicine Indianapolis, IN 46223, USA

Received December 14, 1979. SV40 chromosomes capable of continued RNA synthesis in vitro have been extracted from infected cells in solutions of very low ionic strength (5 mM HEPES, 0.25 mM MgCl2. The RNA made is of high molecular weight, and synthesis is sensitive to {alpha}-amanitin. More RNA is made than from previously described (high salt-extracted) transcription complexes. Transcribing SV4O chromosomes sediment at a rate intermediate between replicating and mature chromosomes, and have a higher ratio of protein to nucleic acid than either. They retain proteins that are lost during exposure to high salt, and might prove valuable in identifying proteins involved in SV40 transcription.


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