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Transformation

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Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...

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Updated: Jul 9, 2026

Electroporation of Mycobacteria

Electroporation of Mycobacteria

Published on: May 23, 2008

An efficient electrotransformation method for three Bacillus species.

Wei Quan¹, Chen-Lu Liu¹, Shao-Xuan Shi¹

¹College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Key Laboratory of Biopesticide Creation and Resource Utilization for Autonomous Region Higher Education Institutions, Hohhot, 010018, China.

Applied Microbiology and Biotechnology

|December 1, 2025

Summary

Optimizing electroporation conditions and using glycine significantly enhanced Bacillus transformation efficiency. This provides an efficient method for genetic manipulation of Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus subtilis.

Keywords:

Bacillus amyloliquefaciens Bacillus subtilis Bacillus velezensis Electroporation Response surface methodology (RSM)Transformation

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Electroporation of Mycobacteria

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Area of Science:

Microbiology
Molecular Biology
Biotechnology

Background:

Bacillus strains are vital in food fermentation and agriculture.
Efficient genetic manipulation requires optimized transformation methods.
Existing methods for Bacillus transformation need improvement for practical applications.

Purpose of the Study:

To optimize electroporation conditions for three Bacillus strains: Bacillus amyloliquefaciens YN-J3, Bacillus velezensis JN-Y2, and Bacillus subtilis S-16.
To identify effective cell wall agents for enhancing Bacillus transformation efficiency.
To establish a robust and efficient system for Bacillus genetic manipulation.

Main Methods:

Orthogonal experiments and response surface analysis were employed to optimize electroporation parameters (OD600, competent cell volume, plasmid concentration, field strength).
Various cell wall agents were screened for their ability to improve bacterial competence.
Transformation efficiency was quantified using CFU/µg DNA.

Main Results:

Optimal transformation parameters were determined for each Bacillus strain, leading to high transformation efficiencies (e.g., 22,198.33 CFU/µg DNA for B.a YN-J3).
Glycine (50 mg/mL) significantly increased transformation efficiency by 40, 36, and 24 times for B.a YN-J3, B.v JN-Y2, and B.s S-16, respectively.
A combined approach of optimized electroporation and glycine treatment yielded a stable and efficient transformation system.

Conclusions:

Optimized electroporation protocols combined with glycine treatment offer a highly efficient method for Bacillus strain genetic manipulation.
This improved transformation system facilitates advancements in the industrial and agricultural applications of Bacillus species.
The study provides a valuable protocol for researchers working with Bacillus transformation.