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Nucleic Acids Research, 1985, Vol. 13, No. 15 5441-5455
© 1985

Articles

Nucleotide sequence and mutational analysis of an immunity repressor gene from Bacillus subtilis temperate phage ø105

Patrick Dhaese, Jozef Seurinck1, Bea De Smet and Marc Van Montagu*

Laboratorium voor Genetica, Rijksuniversiteit Gent K.L. Ledeganckstraat 35, B-9000 Gent, Belgium

*To Whom Correspondence should be addressed

Received June 12, 1985. Accepted July 19, 1985.

We have identified and sequenced a bacteriophage ø105 gene encoding an immunity repressor, the first to be characterized from a temperate phage infecting a Gram-positive host. Using superinfection immunity as an assay for repressor function, the ø105 repressor gene was located within a 740-bpPvuII-HindIII subfragment near the left end of the ø105 EcoRI-F fragment. We show that the repressor is specified by the 5'-proximal coding sequence of a translationally overlapping gene pair, transcribed from right to left on the conventional ø105 map. Comparison of its amino acid sequence (146 residues) with that of a large number of Gram-negative bacterial and phage repressors revealed a putative DNA-binding region between positions 20 and 39. The coding region is preceded by a strong Shine-Dalgarno sequence 5'AAAGGAG 3'. Deletion analysis of the 5'-flanking DNA allowed to identify transcriptional control elements. Their structure, 5' TTGTAT 3' at-35 and 5' TATAAT 3' at -10, strongly suggests that the ø105 repressor gene is transcribed by the major vegetative form of B. subtilis RNA polymerase, as would be expected for an early phage gene.

1Present address: Plant Genetics Systems, Plateaustraat 22, B-9000 Gent, Belgium


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