Analysis of the leuB gene from Corynebacterium glutamicum
Language English Country Germany Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
9720199
DOI
10.1007/s002530051254
Knihovny.cz E-resources
- MeSH
- 3-Isopropylmalate Dehydrogenase MeSH
- Alcohol Oxidoreductases chemistry genetics MeSH
- RNA, Bacterial chemistry MeSH
- Corynebacterium enzymology genetics MeSH
- DNA, Bacterial chemistry MeSH
- DNA Primers chemistry MeSH
- Electrophoresis, Agar Gel MeSH
- Escherichia coli chemistry MeSH
- Leucine chemistry MeSH
- Molecular Sequence Data MeSH
- Molecular Weight MeSH
- Blotting, Northern MeSH
- Plasmids chemistry MeSH
- Promoter Regions, Genetic MeSH
- Gene Expression Regulation, Enzymologic MeSH
- Gene Expression Regulation, Bacterial MeSH
- Restriction Mapping MeSH
- Amino Acid Sequence MeSH
- Base Sequence MeSH
- Sequence Analysis, DNA MeSH
- Sequence Homology, Amino Acid * MeSH
- Blotting, Southern MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 3-Isopropylmalate Dehydrogenase MeSH
- Alcohol Oxidoreductases MeSH
- RNA, Bacterial MeSH
- DNA, Bacterial MeSH
- DNA Primers MeSH
- Leucine MeSH
The leuB gene of Corynebacterium glutamicum was found to be present on a 2.2-kb BamHI-SacI chromosomal fragment which complemented the leuB mutation of Escherichia coli. The activity of 3-isopropylmalate dehydrogenase (EC 1.1.1.85), encoded by the leuB gene, was significantly increased in C. glutamicum cells harbouring a plasmid containing the 2.2-kb fragment. The nucleotide sequence of the C. glutamicum leuB coding region (an open reading frame, ORF, of 1020 bp encoding a polypeptide of 340 amino acids with M(r) of 36 144) was determined. The deduced amino acid sequence of the product of this ORF is highly homologous to those of 3-isopropylmalate dehydrogenases from three species of mycobacteria. The transcriptional start site of the leuB gene was localized 35 bp upstream of its translational start; a functional terminator was detected in the 3' flanking region. Northern hybridization analysis showed that the C. glutamicum leuB gene is transcribed as a single monocistronic RNA (approximately 1.2 kb in size). Activity of the leuB promoter was significantly reduced when leucine was present in the growth medium. This suggests the negative regulation of the leuB expression on the transcriptional level in C. glutamicum cells.
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General and molecular microbiology and microbial genetics in the IM CAS
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