Sequence and expression of an isocitrate dehydrogenase-encoding gene from a polycyclic aromatic hydrocarbon oxidizer, Sphingomonas yanoikuyae B1.
An 18.5-kb DNA fragment was cloned from Sphingomonas yanoikuyae (Sy) B1 (previously Beijerinckia B1). Analysis of a 4.3-kb sequence revealed an isocitrate dehydrogenase (Idh)-encoding gene (idhA), an unidentified open reading frame (ORF) and a partial glucosamine synthetase-encoding ORF (glmS). As in a number of bacteria, Tn7 insertion was found specifically at a site past the stop codon of glmS. The predicted 406-amino-acid sequence of IdhA shows, for the first time, an extensive sequence identity (66%) with an eukaryotic NADP+-specific Idh. The idhA gene was expressed in Escherichia coli. Identical restriction fragments carrying idhA were found in B1, Sy IFO15102 and Sy Q1 (formerly S. paucimobilis Q1), indicating a well-conserved idh gene.
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Biotechnology Research Institute, National Research Council of Canada, Québec, Canada.
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D005945:Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing) / Q000235:genetics; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology* / Q000235:genetics; D007521:Isocitrate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000235:genetics; D016366:Open Reading Frames; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015183:Restriction Mapping; D017421:Sequence Analysis; D012689:Sequence Homology, Nucleic Acid
D004251:DNA Transposable Elements; D011084:Polycyclic Aromatic Hydrocarbons; D012336:RNA, Ribosomal, 16S; D007521:Isocitrate Dehydrogenase; D005945:Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing)