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Structural and functional characterization of IS1358 from Vibrio cholerae
S Dumontier1, P Trieu-Cuot, P Berche
1INSERM U.411, Laboratoire de Microbiologie, Faculté de Médecine Necker- Enfants Malades, 75730 Paris Cedex 15, France.
Journal of Bacteriology
|November 26, 1998
Summary
The insertion sequence IS1358 is present in multiple Vibrio cholerae serovars, suggesting its role in the evolution of cholera strains like the O139 epidemic serovar. This element
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- The emergence of Vibrio cholerae serovar O139 as a new epidemic strain involved genetic changes from the pandemic serovar O1.
- A specific 40-kb DNA fragment replaced a 22-kb region in serovar O1 to form serovar O139.
- This O139-specific DNA fragment contains the insertion sequence IS1358.
Purpose of the Study:
- To investigate the distribution of the IS1358 insertion sequence in various Vibrio cholerae serovars.
- To analyze the genetic characteristics and functionality of the IS1358 element.
- To understand the potential role of IS1358 in the genetic diversity and evolution of Vibrio cholerae.
Main Methods:
- Southern blot analysis was used to detect the presence of IS1358 in different Vibrio cholerae serovars.
- Cloning and sequencing were performed on IS1358 copies from V. cholerae O22 and O155.
- A transposon carrying a kanamycin resistance gene flanked by IS1358 was constructed to study transposition.
- Transposition experiments were conducted in Escherichia coli using a conjugative plasmid.
Main Results:
- IS1358 was detected in Vibrio cholerae serovars O1, O2, O22, O139, and O155, but not in O15, O39, and O141.
- Multiple copies of IS1358 were found in serovars O2, O22, and O155.
- Sequences of IS1358 from O22 and O155 were nearly identical to those from O1 and O139.
- IS1358 demonstrated transposition activity, duplicating 10 bp at the insertion site.
Conclusions:
- The IS1358 insertion sequence is widely distributed across various Vibrio cholerae serovars.
- IS1358 exhibits functional transposition capabilities.
- The presence and mobility of IS1358 may contribute to the genetic variability and evolution of Vibrio cholerae, including the emergence of epidemic strains.