amyP, a reporter gene to study strain degeneration inClostridium acetobutylicumATCC 824
Open Access
- 1 April 2002
- journal article
- research article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 210 (1) , 93-98
- https://doi.org/10.1111/j.1574-6968.2002.tb11165.x
Abstract
Clostridium acetobutylicum produces an extracellular α-amylase when grown on glucose as the sole carbon source. This enzyme was previously characterized from a biochemical point of view but its encoding gene was never identified. The 2283-bp amyP gene encodes a 83 013-Da mature protein with an N-terminal domain that exhibits strong identity to the family 13 glycosyl hydrolases such as the Bacillusα-amylases. Transcriptional analysis revealed that amyP is transcribed in solventogenic but not in acidogenic chemostat cultures. These results are in agreement with the extracellular α-amylase activities indicating that the expression of amyP is regulated at the transcriptional level. amyP is located on the pSOL1 megaplasmid that carries all the genes involved in the final steps of solvent formation. Degeneration of C. acetobutylicum has been associated to the loss of pSOL1. We demonstrate here that amyP can be used as a reporter system to quantitatively follow this phenomenon.Keywords
This publication has 20 references indexed in Scilit:
- Genome Sequence and Comparative Analysis of the Solvent-Producing BacteriumClostridium acetobutylicumJournal of Bacteriology, 2001
- Molecular characterization of the α-amylase genes of Lactobacillus plantarum A6 and Lactobacillus amylovorus reveals an unusual 3′ end structure with direct tandem repeats and suggests a common evolutionary originGene, 1997
- Solvent-forming genes in clostridiaNature, 1996
- Regulation of carbon and electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH on mixtures of glucose and glycerolJournal of Bacteriology, 1994
- Cloning, sequencing, and molecular analysis of the sol operon of Clostridium acetobutylicum, a chromosomal locus involved in solventogenesisJournal of Bacteriology, 1993
- Wide diversity of genome size among different strains of Clostridium acetobutylicumJournal of General Microbiology, 1993
- Cloning, sequencing, and molecular analysis of the acetoacetate decarboxylase gene region from Clostridium acetobutylicumJournal of Bacteriology, 1990
- Molecular cloning, nucleotide sequencing, and expression of the Bacillus subtilis (natto) IAM1212 alpha-amylase gene, which encodes an alpha-amylase structurally similar to but enzymatically distinct from that of B. subtilis 2633Journal of Bacteriology, 1990
- Nucleotide sequence of the amylase gene fromBacillus subtilisNucleic Acids Research, 1983
- REGULATORY SEQUENCES INVOLVED IN THE PROMOTION AND TERMINATION OF RNA TRANSCRIPTIONAnnual Review of Genetics, 1979