发起人
合成生物学
塔塔盒子
计算生物学
金门
生物
抄写(语言学)
DNA结合位点
DNA
遗传学
表观遗传学
基因
基因表达
工程类
土木工程
哲学
跨度(工程)
语言学
作者
Tom Schreiber,Alain Tissier
出处
期刊:Methods in Enzymology
日期:2016-01-01
卷期号:: 361-378
被引量:9
标识
DOI:10.1016/bs.mie.2016.03.004
摘要
The discovery of proteins with programmable DNA-binding specificities triggered a whole array of applications in synthetic biology, including genome editing, regulation of transcription, and epigenetic modifications. Among those, transcription activator-like effectors (TALEs) due to their natural function as transcription regulators, are especially well-suited for the development of orthogonal systems for the control of gene expression. We describe here the construction and testing of libraries of synthetic TALE-activated promoters which are under the control of a single TALE with a given DNA-binding specificity. These libraries consist of a fixed DNA-binding element for the TALE, a TATA box, and variable sequences of 19 bases upstream and 43 bases downstream of the DNA-binding element. These libraries were cloned using a Golden Gate cloning strategy making them usable as standard parts in a modular cloning system. The broad range of promoter activities detected and the versatility of these promoter libraries make them valuable tools for applications in the fine-tuning of expression in metabolic engineering projects or in the design and implementation of regulatory circuits.
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