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ISCce1 and ISCce2, two novel insertion sequences in Clostridium cellulolyticum
Hédia Maamar1, Pascale de Philip, Jean-Pierre Bélaich
1Laboratoire de Bioénergétique et Ingénierie des Protéines, UPR 9036-CNRS, 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Journal of Bacteriology
|January 21, 2003
Summary
Two novel insertion sequences, ISCce1 and ISCce2, disrupt the cipC gene in Clostridium cellulolyticum mutants. These mobile genetic elements, ISCce1 and ISCce2, exhibit characteristics of transposases and appear specific to this bacterial strain.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Spontaneous mutations in Clostridium cellulolyticum can lead to disruptions in essential genes.
- The cipC gene is crucial for cellulose degradation in this species.
Purpose of the Study:
- To identify and characterize novel insertion sequences responsible for disrupting the cipC gene.
- To investigate the nature and potential mobility of these insertion sequences.
Main Methods:
- Analysis of spontaneous mutants of Clostridium cellulolyticum.
- DNA sequencing to identify insertion sequences.
- Open reading frame analysis and protein homology searches.
- Southern blot analysis for genomic copy number assessment.
Main Results:
- Discovery of two new insertion sequences, ISCce1 (1,292 bp) and ISCce2 (1,359 bp).
- ISCce1 and ISCce2 disrupt the cipC gene, encoding putative transposases with similarities to IS481, IS3, and IS256 families.
- Both elements generate 8-bp direct repeats upon transposition.
- ISCce1 is highly abundant (≥20 copies) in the genome, and ISCce2 is often found within ISCce1.
- These sequences appear specific to the studied Clostridium cellulolyticum strain.
Conclusions:
- ISCce1 and ISCce2 are novel mobile genetic elements in Clostridium cellulolyticum.
- Their insertion into the cipC gene contributes to spontaneous mutations.
- The characteristics suggest they are functional transposons with strain-specific prevalence.