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微调 LightOn 光控转基因表达系统。

Fine tuning the LightOn light-switchable transgene expression system.

机构信息

Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, School of Pharmacy, East China University of Science and Technology, Mei Long Road, Shanghai 200237, China.

出版信息

Biochem Biophys Res Commun. 2013 Oct 25;440(3):419-23. doi: 10.1016/j.bbrc.2013.09.092. Epub 2013 Oct 1.

Abstract

Spatiotemporal control of transgene expression in living cells provides new opportunities for the characterization of gene function in complex biological processes. We previously reported a synthetic, light-switchable transgene expression system called LightOn that can be used to control gene expression using blue light. In the present study, we modified the different promoter segments of the light switchable transcription factor GAVPO and the target gene, and assayed their effects on protein expression under dark or light conditions. The results showed that the LightOn system maintained its high on/off ratio under most modifications, but its induction efficiency and background gene expression level can be fine-tuned by modifying the core promoter, the UASG sequence number, the length of the spacer between UASG and the core promoter of the target protein, and the expression level of the GAVPO transcription factor. Thus, the LightOn gene expression system can be adapted to a large range of applications according to the requirements of the background and the induced gene expression.

摘要

活细胞中转基因表达的时空控制为研究基因在复杂生物过程中的功能提供了新的机会。我们之前报道了一种名为 LightOn 的合成、光控转基因表达系统,可用于使用蓝光控制基因表达。在本研究中,我们对光开关转录因子 GAVPO 和目的基因的不同启动子片段进行了修饰,并在暗或光条件下检测了它们对蛋白质表达的影响。结果表明,LightOn 系统在大多数修饰下仍保持较高的开关比,但通过修饰核心启动子、UASG 序列数、UASG 与靶蛋白核心启动子之间的间隔长度以及 GAVPO 转录因子的表达水平,可以精细调整其诱导效率和背景基因表达水平。因此,根据背景和诱导基因表达的要求,LightOn 基因表达系统可以适应广泛的应用。

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