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Nat Biotechnol. 2006 Aug;24(8):1027-32. Epub 2006 Jul 16.

Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes.

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  • 1Department of Chemical Engineering, University of California-Berkeley, Berkeley, CA 94720-1462, USA.

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.

PMID:
16845378
[PubMed - indexed for MEDLINE]
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