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Master thesis

La réparation des cassures double brin d'ADN par NHEJ peut-elle mener à l’intégration d’ADN exogène chez Streptomyces ambofaciens ?

Abstract : Streptomyces bacteria are a model for studying genomic plasticity and chromosomal rearrangements. The Strada team at the Dynamic laboratory wishes to understand the mechanisms involved in this genetic plasticity, by studying the repair of DNA double-stranded breaks (DSB). The team recently demonstrated that in Streptomyces ambofaciens, Non Homologous End Joining (NHEJ), which is inherently mutagenic, allowed the ectopic insertion of endogenous DNA at the site of repair of these breaks. The objective of the internship was to study the involvement of NHEJ in the integration of exogenous DNA in S. ambofaciens. To do this, a conjugation strategy between two S. ambofaciens allowing the transfer of exogenous chromosomal DNA in a receptor bacterium was developed. The strategy was to induce a targeted DSB in the receptor genome using the constitutive expression of a meganuclease, and to determine whether its repair enabled the integration of exogenous DNA. This work demonstrated the transfer and integration of a chromosomal marker into the genome of the receptor at high frequency. The genetic tool allowing the transfer of the meganuclease into the receptor genome during conjugation was validated. The perspective is now to analyse all the repair events at the target DSB in order to screen putative integration of exogenous DNA by NHEJ.
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Master thesis
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https://hal.univ-lorraine.fr/hal-03376603
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Submitted on : Wednesday, October 13, 2021 - 3:48:49 PM
Last modification on : Saturday, April 30, 2022 - 4:02:56 PM

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  • HAL Id : hal-03376603, version 1

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Louise Rostan. La réparation des cassures double brin d'ADN par NHEJ peut-elle mener à l’intégration d’ADN exogène chez Streptomyces ambofaciens ?. Microbiologie et Parasitologie. 2020. ⟨hal-03376603⟩

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