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© Melanie Blokesch, EPFL
Flagellated Vibrio cholerae
Publication : Nucleic acids research

The synthetic integron: an in vivo genetic shuffling device

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nucleic acids research - 09 Jun 2010

Bikard D, Julié-Galau S, Cambray G, Mazel D

Link to Pubmed [PMID] – 20534632

Nucleic Acids Res. 2010 Aug;38(15):e153

As the field of synthetic biology expands, strategies and tools for the rapid construction of new biochemical pathways will become increasingly valuable. Purely rational design of complex biological pathways is inherently limited by the current state of our knowledge. Selection of optimal arrangements of genetic elements from randomized libraries may well be a useful approach for successful engineering. Here, we propose the construction and optimization of metabolic pathways using the inherent gene shuffling activity of a natural bacterial site-specific recombination system, the integron. As a proof of principle, we constructed and optimized a functional tryptophan biosynthetic operon in Escherichia coli. The trpA-E genes along with ‘regulatory’ elements were delivered as individual recombination cassettes in a synthetic integron platform. Integrase-mediated recombination generated thousands of genetic combinations overnight. We were able to isolate a large number of arrangements displaying varying fitness and tryptophan production capacities. Several assemblages required as many as six recombination events and produced as much as 11-fold more tryptophan than the natural gene order in the same context.

http://www.ncbi.nlm.nih.gov/pubmed/20534632