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Updated: Jul 12, 2025

Electroporation of Mycobacteria
Published on: May 23, 2008
Altering under-represented DNA sequences elevates bacterial transformation efficiency
Shuai Hu1, Stefani Giacopazzi1, Ryan Modlin1,2
1Department of Microbiology and Environmental Toxicology, University of California, Santa Cruz, California, USA.
Importance:
Manipulating the genomes of bacteria is critical to many fields. Such manipulations are made by genetic engineering, which often requires new pieces of DNA to be added to the genome. Bacteria have robust systems for identifying and degrading new DNA, some of which rely on restriction enzymes. These enzymes cut DNA at specific sequences. We identified a set of DNA sequences that are missing normally from a bacterium's genome, more than would be expected by chance. Eliminating these sequences from a new piece of DNA allowed it to be incorporated into the bacterial genome at a higher frequency than new DNA containing the sequences. Removing such sequences appears to allow the new DNA to fly under the bacterial radar in "stealth" mode. This transformation improvement approach is straightforward to apply and likely broadly applicable.
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