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Genome-scale promoter engineering by coselection MAGE.

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.

SUBMITTER: Wang HH

PROVIDER: S-EPMC3428217 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature

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Genome-scale promoter engineering by coselection MAGE.

Wang Harris H HH Kim Hwangbeom H Cong Le L Jeong Jaehwan J Bang Duhee D Church George M GM

Nature methods 20120408 6

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-functi ...[more]

PMID: 22484848

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