1. Cloning and sequence analysis of the genes encoding phoshotransbutyrylase and butyrate kinase from Clostridium acetobutylicum NCIMB 8052
- Author
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Nigel P. Minton, John D. Oultram, Michael J. Elmore, and lan D. Burr
- Subjects
DNA, Bacterial ,Butyrate kinase ,Clostridium acetobutylicum ,Sequence analysis ,Recombinant Fusion Proteins ,DNA Mutational Analysis ,Molecular Sequence Data ,Restriction Mapping ,Open Reading Frames ,Phosphate Acetyltransferase ,Plasmid ,Species Specificity ,Escherichia coli ,Genetics ,Amino Acid Sequence ,Cloning, Molecular ,ORFS ,Clostridium ,Dihydropteroate Synthase ,Base Sequence ,Sequence Homology, Amino Acid ,biology ,Phosphotransferases ,Nucleic acid sequence ,General Medicine ,Phosphotransferases (Carboxyl Group Acceptor) ,biology.organism_classification ,Molecular biology ,Complementation ,Open reading frame ,Biochemistry ,Genes, Bacterial ,Bacillus subtilis - Abstract
An 8.1-kb fragment of chromosomal DNA from Clostridium acetobutylicum NCIMB 8052 (formerly NCIB 8052) has been cloned into plasmid pAT153 and shown to allow the growth of Escherichia coli LJ32 (F + ato C2 c ato D32 fadR ) on butyrate as the sole source of carbon and energy. Deletion analysis delineated a 3.9-kb subfragment capable of complementation. The nucleotide sequence of this fragment was determined and it was shown to encode three complete, and two incomplete open reading frames (ORFs). Based on enzymic studies of recombinant clones, two of these ORFs were shown to encode phosphotransbutyrylase and butyrate kinase. The above enzymes are involved in the acidogenic phase of fermentation in C. acetobutylicum . The fragment also carries an incomplete ORF encoding a polypeptide exhibiting substantial similarity to dihydropteroate synthase.
- Published
- 1993
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