Searching for a mechanistic modulator of Tn7 transposition

Project Type: Master

Background:
The classical Tn7 transposon and recently discovered Tn7-CRISPR-Cas elements have been repurposed for development of a genomic DNA integration tool to engineer bacterial chromosomes in a specific manner: integration into the attTn7 site immediately downstream of the glmS gene for the former and into an RNA-guided site for the latter. In those applications, the Tn7 transposition follows a cut-and-paste mechanism, that allows a DNA fragment flanked by the Tn7 right and left ends to be excised from a donor plasmid and integrated into the target site. Nonetheless, the system can be modulated by the presence of a conjugative plasmid to switch from a cut-and-paste mechanism to a replicative manner, in which two copies of the DNA insert and/or a larger DNA fragment can be integrated into a target site, further expanding a potential use of the Tn7 transposon and Tn7-CRISPR-Cas elements. In this project, the candidate will identify the gene/protein encoded by the conjugative plasmid that is able to switch the mechanism of the Tn7 transposition and incorporate the discovered modulator into a new Tn7-mediated DNA integration tool.

Types of projects:
Development of experimental assays to screen a modulator of Tn7 transposition
Development of a replicative Tn7-mediated integration tool
Development of an Tn7-based integrative-conjugative biosensor

Techniques covered:
Molecular cloning
Functional genetics
PCR
TraDIS
DNA Sequencing (Sanger and/or Illumina short-read sequencing)
Conjugation assays

For more information contact:
Assistant professor Joseph Nesme (joseph.nesme@bio.ku.dk)
Professor Søren Sørensen (sjs@bio.ku.dk)

READ MORE AT: https://www1.bio.ku.dk/english/research/microbiology/

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