Broad Purpose Vector for Site-Directed Insertional Mutagenesis in Bifidobacterium breve.

ABSTRACT: Members of the genus Bifidobacterium are notoriously recalcitrant to genetic manipulation due to their extensive and variable repertoire of Restriction-Modification (R-M) systems. Non-replicating plasmids are currently employed to achieve insertional mutagenesis in Bifidobacterium. One of the limitations of using such insertion vectors is the presence within their sequence of various restriction sites, making them sensitive to the activity of endogenous restriction endonucleases encoded by the target strain. For this reason, vectors have been developed with the aim of methylating and protecting the vector using a methylase-positive Escherichia coli strain, in some cases containing a cloned bifidobacterial methylase. Here, we present a mutagenesis approach based on a mo

SUBMITTER: Hoedt EC 

PROVIDER: S-EPMC8021953 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature