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The nuclear DNA of HeLa cells can now be isolated unbroken and supercoiled. Using DNA gyrase and the untwisting enzyme, we have prepared an allomorphic series of templates derived from this nuclear DNA, and also from the circular DNA of the bacterial virus, PM2. We have then transcribed these templates using 2 different RNA polymerases--from wheat germ and Escherichia coli. Relaxed DNA is transcribed slowly by both polymerases. Supertwisting the naturally-supercoiled templates with gyrase slightly inhibits transcription by the bacterial polymerase but stimulates dramatically transcription by RNA polymerase II from wheat germ.

Type

Journal article

Journal

Nucleic Acids Res

Publication Date

25/02/1980

Volume

8

Pages

845 - 854

Keywords

Animals, DNA Topoisomerases, Type I, DNA Topoisomerases, Type II, DNA, Single-Stranded, DNA, Superhelical, DNA, Viral, DNA-Directed RNA Polymerases, Escherichia coli, HeLa Cells, Humans, Rats, Transcription, Genetic, Triticum