Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions.
Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions.
复制标题
DOI：
10.3389/fcimb.2017.00475
复制
发表时间：
2017
影响因子：
5.7
通讯作者：
Steele-Mortimer O
中科院分区：
文献类型：
--
作者：
Cooper KG;Chong A;Starr T;Finn CE;Steele-Mortimer O
在这里，我们描述了使用合成的遗传元素来提高沙门氏菌中表型蛋白生产的可预测性和可调性。我们采用了多管齐下的方法，将一系列强度可变的合成启动子与合成转录终止子结合起来，并改变了质粒拷贝数。这产生了一系列的质粒，在很大的动态范围内驱动荧光蛋白和内源蛋白的统一产生。我们描述了几个例子，其中该系统被用来微调沙门氏菌的结构性表达，提供了一种有效的手段来滴定蛋白质生产的毒性影响。
Here we describe the use of synthetic genetic elements to improve the predictability and tunability of episomal protein production in Salmonella. 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 Salmonella, providing an efficient means to titrate out toxic effects of protein production.