Transcription in Vitro of φ 29 DNA and EcoRI Fragments by Bacillus subtilis RNA Polymerase

Abstract

EcoRI fragments A, B and C produced from linear [phage] .PHI.29 DNA, but not D or E fragments, are transcribed by purified B. subtilis RNA polymerase [EC 2.7.7.6]. The transcription of fragments A and C is initiated preferentially with GTP and to a lesser extent with ATP; the reverse happens in the case of fragment B. The dinucleotides GpU and GpA, respectively, compete specifically with the incorporation of [.gamma.-32P]GTP directed by fragments A and C. The RNA synthesized in vitro by purified B. subtilis RNA polymerase is highly asymmetric. Most of the RNA synthesis directed by fragments A and C is early RNA. Most of the RNA produced by fragment B is anti-late-RNA. Addition of crude extracts inhibit the transcription of fragment B but not that of fragments A and C.