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Nucleic Acids Research, 1982, Vol. 10, No. 13 3905-3919
© 1982

MOLECULAR BIOLOGY

Transcriptional analyses of the Bacillus licheniformis penP gene

Jane R. McLaughlin, Shen-Yung Chang and Shing Chang

Cetus Corporation 600 Bancroft Way, Berkeley, CA 94710, USA

Received March 22, 1982. Revised May 28, 1982. Accepted May 28, 1982.

We report the promoter structure of the Bacillus licheniformis 749/C penicillinase (penP) gene. The transcript encoding the penicillinase gene was identified by in vitro run-off transcription using both E. coli RNA polymerase and B. subtilis RNA polymerase. Utilization of this promoter in linearized DNA by the B. subtilis RNA polymerase showed extreme sensitivity to ionic strength. The 5' sequence of the penP mRNA was determined using enzymatic sequencing (1). Holoenzymes from E. coli and B. subtilis (Eo55) initiate penP RNA synthesis at the same site. Alignment of this RNA sequence with the reported DNA sequence of the penP gene (2, 3) revealed the "–35 region" and "Pribnow box" sequences that are recognized as 5'TTGCAT and 5'AATACT, respectively. Potential secondary structure within the promoter exists which may play a role in the expression of the penicillinase gene. The location of the penP promoter was further confirmed by molecular cloning of a DNA fragment containing the expected promoter sequence into a promoter-fishing vector useful for monitoring promoter activities in both E. coli and B. subtilis.


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