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RNA Polymerase binding sites and transcription map of the DNA of Bacillus subtilis phage φ29

AuthorsInciarte, M. R.; Corral, Javier; Viñuela, Eladio; Salas, Margarita ; Sogo, José M.
Issue Date5-Feb-1979
CitationJournal of Molecular Biology 127(4): 411-436 (1979)
AbstractThe Bacillus subtilis RNA polymerase bound to phage φ29 DNA has been visualized by electron microscopy. Seven specific binding sites have been observed at 1.7(±0.4)†, 25.5(±0.5), 26.7(±0.4), 59.4(±1.2), 79.3(±0.9), 91.3(±0.6) and 99sd3(± 0.4) DNA length units (one unit is equal to 1% of the length of φ29 DNA). Three of the sites are located in fragment EcoRI A, two in fragment EcoRI B and two in fragment EcoRI C. The same binding sites are seen whether proteinase K-treated φ29 DNA or protein p3-containing φ29 DNA is used. By hybridization of early or late φ29 induced RNA, at saturation, to the separated strands of uniformly labelled φ29[32P]DNA restriction fragments, which cover more than 95% of the genome, we have determined the extent of the in vivo early transcription from the L strand of fragments EcoRI A, B and D and HpaII C and that of late transcription from the complementary strands (H strand). We have shown the existence of symmetric transcription within most of fragment EcoRI B and in about one third of fragment EcoRI D. We have also found that sus mutants in cistron 4 do not induce the synthesis of late RNA suggesting that protein p4 is involved in the control of late transcription.
Publisher version (URL)http://dx.doi.org/10.1016/0022-2836(79)90230-4
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