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Precise excision of bacteriophage Mu DNA

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Abstract:

The temperate bacteriophage Mu is a transposable element that can integrate randomly into bacterial DNA, thereby creating mutations. Mutants due to an integrated Mu prophage do not give rise to revertants, as if Mu, unlike other transposable elements, were unable to excise precisely. In the present work, starting with a lacZ::Muc62(Ts) strain unable to form Lac+ colonies, we cloned a lacZ+ gene in vivo on a mini-Mu plasmid, under conditions of prophage induction. In all lac+ plasmids recovered, the wild-type sequence was restored in the region where the Mu prophage had been integrated. The recovery of lacZ+ genes shows that precise excision of Mu does indeed take place; the absence of Lac+ colonies suggests that precise excision events are systematically associated with loss of colony-forming ability.Key words: transposable element, Escherichia coli, mini-Muduction.

Le bactériophage tempéré Mu est un élément transposable qui s'intègre au hasard dans l'ADN bactérien, créant ainsi des mutations. Les mutants résultant d'un prophage Mu intégré ne réversent pas, comme si Mu, à la différence d'autres éléments transposables, était incapable de s'exciser précisément. Dans le présent travail, à partir d'une souche lacZ::Muc62(Ts) incapable de donner des colonies Lac+, nous avons cloné un gène lacZ+ in vivo sur un plasmide mini-Mu, dans des conditions d'induction du prophage. Dans tous les plasmides lac+ récupérés, la séquence du gène sauvage a été restaurée dans la région où le prophage Mu avait été intégré. L'obtention de gènes lacZ+ démontre que l'excision précise de Mu a effectivement lieu; l'absence de colonies Lac+ suggère que les événements d'excision précise sont systématiquement associés à la perte de la capacité à former une colonie.Mots clés : élément transposable, Escherichia coli, mini-Muduction.

Keywords: Escherichia coli; mini-Muduction; transposable element; élément transposable

Document Type: Research Article

Publication date: August 1, 2001

More about this publication?
  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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