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Nat Commun. 2016 Oct 26;7:13215. doi: 10.1038/ncomms13215.

A robust gene-stacking method utilizing yeast assembly for plant synthetic biology.

Shih PM1,2, Vuu K1,2, Mansoori N1,2, Ayad L1,2, Louie KB3,4, Bowen BP3,4, Northen TR1,3,4, Loqué D1,2,5.

Author information

1
Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California 94608, USA.
2
Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, Berkeley, California 94720, USA.
3
Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, Berkeley, California 94720, USA.
4
Joint Genome Institute, 2800 Mitchell Drive, Walnut Creek, California 94598, USA.
5
Université Claude Bernard Lyon 1, INSA de Lyon, CNRS, UMR5240, Microbiologie, Adaptation et Pathogénie, 10 rue Raphaël Dubois, F-69622 Villeurbanne, France.

Abstract

The advent and growth of synthetic biology has demonstrated its potential as a promising avenue of research to address many societal needs. However, plant synthetic biology efforts have been hampered by a dearth of DNA part libraries, versatile transformation vectors and efficient assembly strategies. Here, we describe a versatile system (named jStack) utilizing yeast homologous recombination to efficiently assemble DNA into plant transformation vectors. We demonstrate how this method can facilitate pathway engineering of molecules of pharmaceutical interest, production of potential biofuels and shuffling of disease-resistance traits between crop species. Our approach provides a powerful alternative to conventional strategies for stacking genes and traits to address many impending environmental and agricultural challenges.

PMID:
27782150
PMCID:
PMC5095168
DOI:
10.1038/ncomms13215
[Indexed for MEDLINE]
Free PMC Article

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