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Nucleotide sequence of the penicillinase repressor gene penI of Bacillus licheniformis and regulation of penP and penI by the repressor
- Source :
- Journal of Bacteriology. 168:1128-1132
- Publication Year :
- 1986
- Publisher :
- American Society for Microbiology, 1986.
-
Abstract
- Bacillus licheniformis penicillinase genes, penP and penI, are coded on a 4.2-kilobase EcoRI fragment of pTTE21 (T. Imanaka, T. Tanaka, H. Tsunekawa, and S. Aiba, J. Bacteriol. 147:776-186, 1981). The EcoRI fragment was subcloned in a low-copy-number plasmid pTB522 in Bacillus subtilis. B. subtilis carrying the recombinant plasmid pPTB60 (Tcr penP+ penI+) was chemically mutagenized. Of about 150,000 colonies, two penI(Ts) mutant plasmids, pPTB60D13 and pPTB60E24, were screened by the plate assay at 30 and 48 degrees C for penicillinase. By constructing recombinant plasmids between wild-type and mutant plasmids, the mutation points were shown to be located in a 1.7-kilobase EcoRI-PstI fragment. The EcoRI-PstI fragments of the wild-type plasmid and two mutant plasmids were sequenced. A large open reading frame, composed of 384 bases and 128 amino acid residues (molecular weight, 14,983), was found. Since the mutation points were located at different positions in the protein coding region (Ala to Val for pPTB60D13 and Pro to Leu for pPTB60E24), the coding region was concluded to be the penI gene. A Shine-Dalgarno sequence was found 7 bases upstream from the translation start site (ATG). A probable promoter sequence which is very similar to the consensus sequence was also found upstream of the penP promoter, but in the opposite direction. A consensus twofold symmetric sequence (AAAGTATTA CATATGTAAGNTTT) which might have been used as a repressor binding region was found downstream and in the midst of the penP promoter and also downstream of the penI promoter. The regulation of penP and penI by the repressor is discussed.
- Subjects :
- Mutant
EcoRI
Repressor
Bacillus
Bacillus subtilis
Microbiology
Plasmid
Bacterial Proteins
Consensus sequence
Amino Acid Sequence
Promoter Regions, Genetic
Molecular Biology
Peptide sequence
Genetics
Base Sequence
biology
Nucleic acid sequence
Penicillinase
biology.organism_classification
Molecular biology
Recombinant Proteins
Repressor Proteins
Gene Expression Regulation
Genes, Bacterial
biology.protein
Research Article
Plasmids
Transcription Factors
Subjects
Details
- ISSN :
- 10985530 and 00219193
- Volume :
- 168
- Database :
- OpenAIRE
- Journal :
- Journal of Bacteriology
- Accession number :
- edsair.doi.dedup.....36fd236b36e22396f907bad3e86d7765
- Full Text :
- https://doi.org/10.1128/jb.168.3.1128-1132.1986