利用转录后调控元件文库对合成操纵子进行优化。
Optimization of synthetic operons using libraries of post-transcriptional regulatory elements.
作者信息
Agnew Daniel E, Pfleger Brian F
机构信息
Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI, USA.
出版信息
Methods Mol Biol. 2011;765:99-111. doi: 10.1007/978-1-61779-197-0_7.
Abstract
Constructing polycistronic operons is an advantageous strategy for coordinating the expression of -multiple genes in a prokaryotic host. Unfortunately, a basic construct consisting of an inducible promoter and genes cloned in series does not generally lead to optimal results. Here, a combinatorial approach for tuning relative gene expression in operons is presented. The method constructs libraries of post--transcriptional regulatory elements that can be cloned into the noncoding sequence between genes. Libraries can be screened to identify sequences that optimize expression of metabolic pathways, multisubunit proteins, or other situations where precise stoichiometric ratios of proteins are desired.
摘要
构建多顺反子操纵子是在原核宿主中协调多个基因表达的一种有利策略。不幸的是，由诱导型启动子和串联克隆的基因组成的基本构建体通常不会产生最佳结果。在此，提出了一种用于调节操纵子中相对基因表达的组合方法。该方法构建可克隆到基因间非编码序列中的转录后调控元件文库。可以筛选文库以鉴定优化代谢途径、多亚基蛋白或其他需要精确蛋白质化学计量比的情况的表达的序列。
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