Summary
The DNA of bacteriophage Mu has been transcribed in vitro with the DNA-dependent RNA polymerase (nucleoside-triphosphate: RNA nucleotidyl transferase, E.C.2.7.7.6) from Escherichia coli. The synthesized RNA was analyzed by hybridization to the separated DNA-strands from bacteriophage Mu.
Compared with the in vivo RNA synthesis of bacteriophage Mu, which is almost exclusively on the r-strand, the in vitro RNA synthesis is apparently not very specific, as about 40% of this synthesis takes place on the l-strand.
The in vitro RNA was also hybridized to the separated DNA-strands of various λpMu transducing phages. These λpMu phages contain parts of the early region of Mu, that differ in length. It appears, that in these phages the in vitro RNA synthesis from the early region is largely on the r-strand, as it also is in vivo. This indicates that the in vivo early promoter is also recognized in vitro. Furthermore this transcription appears to terminate after the early genes. This termination is independent of termination factor rho.
Little RNA, that originates from gene C and the late genes, can be detected. This suggests, that the promoter(s) for these genes might not be active in vitro.
The r-strand of the ‘G’β-area is transcribed at a higher frequency than the middle part of the Mu genome. Such an increase of the transcription frequency can be explained by the presence of a RNA synthesis initiation site on the r-strand in the ‘G’β-area.
The extensive transcription on the l-strand shows a gradient from the ‘G’β-end towards the immunity end. This transcription could be initiated on an E. coli promoter, located within the variable end, or on a Mu promoter in the β segment.
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Communicated by J. Schell
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van Meeteren, R., van de Putte, P. Transcription of bacteriophage Mu. Molec. Gen. Genet. 179, 177–183 (1980). https://doi.org/10.1007/BF00268461
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DOI: https://doi.org/10.1007/BF00268461