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PMID: 6287425 Published · ppublish English Journal Article

Transcriptional analyses of the Bacillus licheniformis penP gene.

Nucleic acids research ·Vol. 10 ·No. 13 ·1982-07-10 ·Pages 3905-19

McLaughlin JR, Chang SY, Chang S

Abstract

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 (E sigma 55) initiate penP RNA synthesis at the same site. Alignment of this RNA sequence with the reported DNA sequence of ther 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.

MeSH Terms
Bacillus/enzymology,genetics Bacillus subtilis/enzymology Base Sequence DNA Restriction Enzymes DNA-Directed RNA Polymerases/metabolism Escherichia coli/enzymology Genes, Bacterial Operon Penicillinase/genetics Transcription, Genetic beta-Lactamases/genetics
Chemicals
DNA-Directed RNA Polymerases DNA Restriction Enzymes Penicillinase beta-Lactamases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
McLaughlin J R
Chang S Y
Chang S
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37 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/10.13.3905
Published
1982-07-10
Pages
3905-19
Language
English
Region
England
NLM ID
0411011
PMCID
PMC320767
Subset
IM
Databases
GENBANK
J01544, J01545
External Links
PubMed source DOI: 10.1093/nar/10.13.3905 PMC Full Text View journal details
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