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Optimization of Electroporation Conditions for Introducing Heterologous DNA into . | LitMetric

Optimization of Electroporation Conditions for Introducing Heterologous DNA into .

J Microbiol Biotechnol

Department of Bio-Environmental Chemistry, College of Agricultural and Life Sciences, Chungnam National University, Daejeon 34134, Republic of Korea.

Published: November 2024

AI Article Synopsis

  • The studied strain can grow using both light and chemical energy, making it effective for turning carbon dioxide into valuable products, but it struggles with low transformation efficiency.
  • The researchers improved this efficiency by knocking out a gene that produces a DNA restriction enzyme, which inhibits transformation.
  • They optimized electroporation conditions, leading to a significant increase in transformants—over 7,000 per µg of DNA—making the strain more useful for industrial applications.

Article Abstract

is a strain capable of both photoautotrophic and chemoautotrophic growth, with various metabolic pathways that make it highly suitable for converting carbon dioxide into high value-added products. However, its low transformation efficiency has posed challenges for genetic and metabolic engineering of this strain. In this study, we aimed to increase the transformation efficiency of by deleting the gene coding for an endogenous DNA restriction enzyme that inhibits. We evaluated the effects of growth conditions for making electrocompetent cells and optimized electroporation parameters to be a cuvette width of 0.1 cm, an electric field strength of 30 kV/cm, a resistance of 200 Ω, and a plasmid DNA amount of 0.5 μg, followed by a 24-h recovery period. As a result, we observed over 7,000 transformants per μg of DNA under the optimized electroporation conditions using the strain, which is approximately 10 times higher than that of wild-type under standard bacterial electroporation conditions. These findings are expected to enhance the application of in various industrial fields in the future.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11637821PMC
http://dx.doi.org/10.4014/jmb.2408.08044DOI Listing

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