Nature Biotechnology
- 24, 1027 - 1032 (2006)
Published online: 16 July 2006; | doi:10.1038/nbt1226
Combinatorial engineering of intergenic regions in operons tunes expression of multiple genesBrian F Pfleger1, Douglas J Pitera1, Christina D Smolke1, 4 & Jay D Keasling1, 2, 31
Department of Chemical Engineering, University of California Berkeley, California 94720-1462, USA. 2
Department of Bioengineering, University of California Berkeley, California 94720-1762, USA. 3
Synthetic Biology Department, Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA. 4
Present address: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Correspondence should be addressed to Jay D Keasling keasling@berkeley.edu 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.
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