Related Experiment Video
Updated: Jun 5, 2026

Methods for Electroporation and Transformation Confirmation in Limosilactobacillus reuteri DSM20016
Published on: June 23, 2023
Overcoming the restriction barrier to plasmid transformation and targeted mutagenesis in Bifidobacterium breve
Mary O'Connell Motherway1, Jonathan O'Driscoll, Gerald F Fitzgerald
1Alimentary Pharmabiotic Centre, Department of Microbiology and Department of Food and Nutritional Sciences , National University of Ireland, Cork, Western Road, Cork, Ireland.
Abstract:
In silico analysis of the Bifidobacterium breve UCC2003 genome predicted two distinct loci, which encode three different restriction/modification systems, each comprising a modification methylase and a restriction endonuclease. Based on sequence homology and observed protection against restriction we conclude that the first restriction endonuclease, designated BbrI, is an isoschizomer of BbeI, the second, BbrII, is a neoschizomer of SalI, while the third, BbrIII, is an isoschizomer of PstI. Expression of each of the B. breve UCC2003 methylase-encoding genes in B. breve JCM 7017 established that BbrII and BbrIII are active and restrict incoming DNA. By exploiting knowledge on restriction/modification in B. breve UCC2003 we successfully increased the transformation efficiency to a level that allows the reliable generation of mutants by homologous recombination using a non-replicative plasmid.
Related Concept Videos
Bacterial Transformation
Bacterial Transformation
Antibiotic Selection
Transformation

