Nucleic Acids Research Advance Access originally published online on December 12, 2008
Nucleic Acids Research 2009 37(2):e16; doi:10.1093/nar/gkn991
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Nucleic Acids Research, 2009, Vol. 37, No. 2 e16
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways
1Department of Chemical and Biomolecular Engineering and 2Department of Chemistry, Biochemistry, and Bioengineering, Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
*To whom correspondence should be addressed. Tel: +1 217 333 2631; Fax: +1 217 333 5052; Email: zhao5{at}illinois.edu
Received April 9, 2008. Revised November 24, 2008. Accepted November 25, 2008.
The assembly of large recombinant DNA encoding a whole biochemical pathway or genome represents a significant challenge. Here, we report a new method, DNA assembler, which allows the assembly of an entire biochemical pathway in a single step via in vivo homologous recombination in Saccharomyces cerevisiae. We show that DNA assembler can rapidly assemble a functional D-xylose utilization pathway (
9 kb DNA consisting of three genes), a functional zeaxanthin biosynthesis pathway (11 kb DNA consisting of five genes) and a functional combined D-xylose utilization and zeaxanthin biosynthesis pathway (19 kb consisting of eight genes) with high efficiencies (70–100%) either on a plasmid or on a yeast chromosome. As this new method only requires simple DNA preparation and one-step yeast transformation, it represents a powerful tool in the construction of biochemical pathways for synthetic biology, metabolic engineering and functional genomics studies.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
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