Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes
Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes
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DOI：
10.1038/nbt1226
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发表时间：
2006-08-01
影响因子：
46.9
通讯作者：
Keasling, Jay D.
中科院分区：
文献类型：
--
作者：
Pfleger, Brian F.;Pitera, Douglas J.;Keasling, Jay D.
合成生物学的许多应用需要多个基因的平衡表达。虽然操纵子促进了原核生物和真核生物中多个基因的协调表达，但通过先验设计协调许多决定操纵子中基因相对表达水平的转录后过程仍然是一个挑战。我们描述了一种通过生成可调基因间区（TIGRs）文库、重组各种转录后控制元件和筛选所需相对表达水平来调节操纵子内多个基因表达的方法。TIGRs可以在100倍的范围内改变两个报告基因的相对表达，并在编码异源甲羟戊酸生物合成途径的操纵子中平衡三个基因的表达，导致甲羟戊酸产量增加7倍。该技术可用于优化合成操纵子中多个基因的表达，无论是在原核生物还是真核生物中。
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.