Optimization of Electroporation Conditions for Bacillus pumilus 3–19 Strain

Optimization of Electroporation Conditions for Bacillus pumilus 3–19 Strain

Electroporation is the process of using electrical impulses to create temporary pores in the plasma membrane which, in turn, enables the penetration of nucleic acids into the cytoplasm of a cell. This method is widely used for the rapid and efficient introduction of foreign DNA into a wide range of...

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Bibliographic Details
Published in:BioNanoScience Vol. 12; no. 3; pp. 752 - 756
Main Authors: Danilova, I. V., Rudakova, N. L., Vasilyeva, Y. A., Gilmutdinova, A. I., Diadkina, I. V., Khasanov, D. I., Sharipova, M. R.
Format: Journal Article
Language:English
Published: New York Springer US 01-09-2022
Springer Nature B.V
Subjects:
Bacillus pumilus
Biological and Medical Physics
Biomaterials
Biophysics
Buffers
Cell culture
Cell viability
Circuits and Systems
Cytoplasm
Deoxyribonucleic acid
DNA
Efficiency
Electric field strength
Electric pulses
Electroporation
Engineering
Nanotechnology
Nucleic acids
Optimization
Electroporation
Transformation
Competent cells
Bacterial growth phase
Electric field strength
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Summary:Electroporation is the process of using electrical impulses to create temporary pores in the plasma membrane which, in turn, enables the penetration of nucleic acids into the cytoplasm of a cell. This method is widely used for the rapid and efficient introduction of foreign DNA into a wide range of cells. Cell viability and electrotransfection efficiency depend on various experimental factors, including the impulse form, vector concentration, cell type (density), properties of the electroporation buffer, and the growth phase of the bacterial culture. In this work, we investigated the optimal conditions for the transformation of the Bacillus pumilus 3–19 strain using electroporation. Competent B. pumilus cells were obtained on the 4th hour of culture growth using PEB1 electroporation buffer and SOC1 (Super Optimal broth with Catabolic repressor) medium, while the electric field strength was 12 kV/cm. With these parameters, the transformation efficiency of bacillus cells was 56.3 transformants/μg DNA. Thus, the rational choice of pulsation conditions and buffering compositions is critical for the design of electroporation protocols to maximize the viability and efficiency of electrotransfection.
ISSN:2191-1630
2191-1649
DOI:10.1007/s12668-022-00982-2