Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.
Front Cell Infect Microbiol. 2017 Nov 16;7:475. doi: 10.3389/fcimb.2017.00475. eCollection 2017.
Here we describe the use of synthetic genetic elements to improve the predictability and tunability of episomal protein production in . We used a multi-pronged approach, in which a series of variable-strength synthetic promoters were combined with a synthetic transcriptional terminator, and plasmid copy number variation. This yielded a series of plasmids that drive uniform production of fluorescent and endogenous proteins, over a wide dynamic range. We describe several examples where this system is used to fine-tune constitutive expression in , providing an efficient means to titrate out toxic effects of protein production.
在这里,我们描述了使用合成遗传元件来提高质体蛋白生产的可预测性和可调节性。我们采用了一种多管齐下的方法,其中一系列可变强度的合成启动子与合成转录终止子和质粒拷贝数变化相结合。这产生了一系列质粒,可在很宽的动态范围内均匀地驱动荧光和内源性蛋白的产生。我们描述了几个例子,其中该系统用于微调在中的组成型表达,为滴定蛋白生产的毒性效应提供了一种有效的方法。