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Nucleic Acids Research, 1978, Vol. 5, No. 7 2665-2678
© 1978

Articles

In vitro transcription in E.coli crude lysates prepared on cellophane discs

Svein Valla1 and Bjöm H. Lindqvist2

Institute of Medical Biology, University of Tromsø 9001 Tromsø, Norway

2To whom reprint request should be addressed

Received February 27, 1978.

An in vitro RNA-synthesizing system consisting of gently lysed E. coli cells on cellophane discs is described. The system has been optimalized with respect to total RNA synthesis. Under certain standard conditions DNA dependent RNA polymerase (EC 2.7.7.6 [EC] ) is responsible for the majority of the RNA synthesis. The extensive rifampicin sensitivity of the synthesis indicates that most of the transcripts are initiated in vitro The RNA synthesizing system described here has been developed with the aim of studying phage transcription in vitro We show here that lysates of a P4 infected P2 lysogen support initiation and propagation of transcription from the P2 prophage.

1Present address: Institutt for teknisk biokjemi, Norges tekniske hø 7034 Trondheim, Norway


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