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- Title
Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes.
- Authors
Pfleger, Brian F; Pitera, Douglas J; Smolke, Christina D; Keasling, Jay D
- Abstract
Many applications of synthetic biology require the balanced expression of multiple genes. Although operons facilitate coordinated expression of multiple genes in prokaryotes and eukaryotes, coordinating the many post-transcriptional processes that determine the relative levels of gene expression in operons by a priori design remains a challenge. We describe a method for tuning the expression of multiple genes within operons by generating libraries of tunable intergenic regions (TIGRs), recombining various post-transcriptional control elements and screening for the desired relative expression levels. TIGRs can vary the relative expression of two reporter genes over a 100-fold range and balance expression of three genes in an operon that encodes a heterologous mevalonate biosynthetic pathway, resulting in a sevenfold increase in mevalonate production. This technology should be useful for optimizing the expression of multiple genes in synthetic operons, both in prokaryotes and eukaryotes.
- Publication
Nature biotechnology, 2006, Vol 24, Issue 8, p1027
- ISSN
1087-0156
- Publication type
Journal Article
- DOI
10.1038/nbt1226