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Nat Methods. 2012 Jun;9(6):591-3. doi: 10.1038/nmeth.1971. Epub 2012 Apr 8.

Genome-scale promoter engineering by coselection MAGE.

Author information

  • 1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. harris.wang@wyss.harvard.edu

Abstract

Multiplex automated genome engineering (MAGE) uses short oligonucleotides to scarlessly modify genomes; however, insertions >10 bases are still inefficient but can be improved substantially by selection of highly modified chromosomes. Here we describe 'coselection' MAGE (CoS-MAGE) to optimize biosynthesis of aromatic amino acid derivatives by combinatorially inserting multiple T7 promoters simultaneously into 12 genomic operons. Promoter libraries can be quickly generated to study gain-of-function epistatic interactions in gene networks.

PMID:
22484848
[PubMed - indexed for MEDLINE]
PMCID:
PMC3428217
Free PMC Article

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