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Genome-Scale Promoter Engineering by Coselection MAGE

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2012

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Nature Publishing Group
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Wang, Harris He, Hwangbeom Kim, Le Cong, Jaehwan Jeong, Duhee Bang, and George McDonald Church. 2012. Genome-scale promoter engineering by coselection MAGE. Nature Methods 9(6): 591-593.

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Abstract

Multiplex Automated Genome Engineering (MAGE) employs short oligonucleotides to scarlessly modify genomes. However, insertions of >10 bases are still inefficient, but can be improved substantially by selection of highly modified chromosomes. Here, we describe Co-Selection 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.

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synthetic biology, systems biology, genomics, molecular biology

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