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Efficient transformation of Lactobacillus sake by electroporation
Françoise Berthier1, Monique Zagorec1, Marie Champomier-Vergès1
1Laboratoire de Recherches sur la Viande, Institut National de la Recherche Agronomique, Domaine de Vilvert, 78352 Jouy-en-Josas Cedex, France.
Microbiology (Reading, England)
|March 17, 2021
Summary
This study optimized electroporation for Lactobacillus sake transformation, achieving high efficiencies up to 10^7 transformants per microgram of DNA. The developed method is effective across multiple Lactobacillus sake strains.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- Lactobacillus sake is a significant lactic acid bacterium.
- Efficient genetic manipulation of Lactobacillus sake is crucial for research and industrial applications.
- Electroporation is a common method for bacterial transformation.
Purpose of the Study:
- To develop and optimize an electroporation procedure for efficient transformation of intact Lactobacillus sake cells.
- To identify critical parameters influencing transformation efficiency in Lactobacillus sake.
- To establish a reliable method applicable to various Lactobacillus sake strains.
Main Methods:
- Systematic examination of electroporation parameters, including electrical settings and solution compositions.
- Optimization of washing, electroporation, and storage solutions.
- Investigation of the role of MgCl2 in the expression medium.
- Testing the optimized procedure on multiple Lactobacillus sake strains.
Main Results:
- Identified electrical parameters, solution composition, and MgCl2 presence as critical factors for transformation efficiency.
- Achieved transformation efficiencies up to 10^7 transformants (μg supercoiled DNA)^-1 under optimal conditions.
- Demonstrated consistent transformation efficiencies ranging from 10^4 to 10^7 transformants (μg supercoiled DNA)^-1 across different Lactobacillus sake strains.
Conclusions:
- An optimized electroporation protocol for Lactobacillus sake has been successfully developed.
- The procedure is robust and yields high transformation efficiencies.
- This method facilitates genetic engineering of Lactobacillus sake for various applications.
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