Reference - Detail
RRC ID | 11217 |
---|---|
Author | O'Connell Motherway M, O'Driscoll J, Fitzgerald GF, Van Sinderen D. |
Title | Overcoming the restriction barrier to plasmid transformation and targeted mutagenesis in Bifidobacterium breve UCC2003. |
Journal | Microb Biotechnol |
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. |
Volume | 2(3) |
Pages | 321-32 |
Published | 2009-5-1 |
DOI | 10.1111/j.1751-7915.2008.00071.x |
PMID | 21261927 |
PMC | PMC3815753 |
MeSH | Amino Acid Sequence Bacterial Proteins / chemistry Bacterial Proteins / genetics* Bacterial Proteins / metabolism Bifidobacterium / chemistry Bifidobacterium / enzymology* Bifidobacterium / genetics* DNA Restriction Enzymes / chemistry DNA Restriction Enzymes / genetics* DNA Restriction Enzymes / metabolism Molecular Sequence Data Mutagenesis* Plasmids / genetics* Sequence Alignment Transformation, Bacterial* |
IF | 5.328 |
Times Cited | 73 |
Resource | |
General Microbes | JCM 7017 JCM 7019 |