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Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates
Published on: May 30, 2017
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Natural Transformation in a Classical-Biotype Vibrio cholerae Strain
Cameron J Lloyd1,2, Adrian Mejia-Santana1,2, Triana N Dalia3
1South Texas Center for Emerging Infectious Diseases, University of Texas San Antonio, San Antonio, Texas, USA.
Applied and Environmental Microbiology
|March 13, 2021
Summary
Classical Vibrio cholerae can take up DNA, enabling genetic manipulation. This study identified key regulators (HapR, TfoX, QstR) for natural transformation in classical strains, overcoming previous limitations.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Vibrio cholerae causes cholera pandemics, with El Tor strains currently dominant over classical strains.
- El Tor strains exhibit efficient natural transformation for DNA acquisition, unlike classical strains.
- Natural transformation is crucial for Vibrio cholerae evolution and adaptation.
Purpose of the Study:
- To investigate the genetic basis of natural transformation in classical Vibrio cholerae strains.
- To enable efficient genetic manipulation of classical strains using natural transformation.
- To compare the natural transformation capabilities of classical and El Tor biotypes.
Main Methods:
- Genetic manipulation of the classical strain O395.
- Artificial induction of competence regulators (HapR, TfoX, QstR).
- Development of a plasmid for simultaneous TfoX and QstR expression.
- Application of multiplex genome editing by natural transformation (MuGENT).
Main Results:
- Artificial induction of TfoX and QstR, alongside HapR, enabled efficient DNA uptake in O395.
- Natural transformation in classical strains remains lower than in El Tor strains.
- A developed plasmid facilitated consistent DNA uptake and multiplex genome editing in O395.
Conclusions:
- Classical Vibrio cholerae O395 is functionally capable of natural transformation.
- The study overcomes limitations in classical strain genetic manipulation.
- Lower transformation efficiency suggests classical strains may utilize horizontal gene transfer less effectively.
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