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Approaches for site-directed mutagenesis in bifidobacteria. Flow chart describing the original (inclusive of red pathway) and modified steps (exclusion of red pathway which can consist of either methylase cloning or vector methylation) required for performing site-directed mutagenesis in Bifidobacterium. The improved pFREM28 vector reduces labor and consumable cost as a step for methylation of the E. coli-Bifidobacterium shuttle vector is no longer required.
