Functional analysis of the pBC1 replicon from Bifidobacterium catenulatum L48 View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2007-08-18

AUTHORS

Pablo Álvarez-Martín, Mary O’Connell-Motherway, Douwe van Sinderen, Baltasar Mayo

ABSTRACT

To determine the minimal replicon of pBC1 (a 2.5-kb cryptic plasmid of Bifidobacterium catenulatum L48) and to check the functionality of its identified open reading frames (ORFs) and surrounding sequences, different segments of pBC1 were amplified by polymerase chain reaction (PCR) and cloned into pBif, a replication probe vector for bifidobacteria. The largest fragment tested in this manner encompassed most of the pBC1 sequence, while the shortest just included the repB gene and its immediate upstream sequences. Derivatives were all shown to allow replication in bifidobacteria. Surprisingly, both the transformation frequency and segregational stability in the absence of antibiotic selection decreased with reducing plasmid length. The relative copy number of the constructs (ranging from around 3 to 23 copies per chromosome equivalent, as compared to 30 copies for the original pBC1) was shown to be strain dependent and to decrease with reducing plasmid length. These results suggest that, although not essential, the copG-like and orfX-like genes of pBC1 play important roles in pBC1 replication. Interruption of repB produced a construct incapable of replicating in bifidobacteria. The analysis of pBC1 will allow its use in the construction of general and specific cloning vectors. More... »

PAGES

1395

Journal

TITLE

Applied Microbiology and Biotechnology

ISSUE

6

VOLUME

76

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00253-007-1115-5

    DOI

    http://dx.doi.org/10.1007/s00253-007-1115-5

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1033037658

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/17704917


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