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. Author manuscript; available in PMC: 2014 Mar 15.
Published in final edited form as: Methods. 2012 Mar 23;60(1):81–90. doi: 10.1016/j.ymeth.2012.03.009

Figure 2.

Figure 2

Control of multiple gene expression levels within operons through tunable intergenic regions (TIGR). A library of TIGR sequences was assembled combinatorially from four sets of oligonucleotides using PCR. A megaprimer PCR approach was used to simultaneously place TIGR libraries within the intergenic regions of an operon with promoter (designated P) and terminator (designated T). The various TIGRs caused the mRNA coding regions to be differentially processed and translated, resulting in different levels of the various proteins encoded in the operon. In some TIGRs, RNase E sites (designated E) were incorporated to decouple the stability of the coding regions and thus the production of the corresponding proteins.