Short Synthetic Terminators for Improved Heterologous Gene Expression in Yeast
Short Synthetic Terminators for Improved Heterologous Gene Expression in Yeast
复制标题
DOI：
10.1021/sb5003357
复制
发表时间：
2015-07-01
影响因子：
4.7
通讯作者：
Alper, Hal S.
中科院分区：
文献类型：
--
作者：
Curran, Kathleen A.;Morse, Nicholas J.;Alper, Hal S.
终止子在完成转录过程和影响mRNA半衰期方面都起着重要作用。因此，终止子是在诸如异源基因表达和代谢工程的应用中考虑的重要合成组分。在这里，我们描述了一组短（35-70 bp）的合成终止子，可用于调节酵母中的基因表达。与常用的CYC 1终止子相比，这些合成终止子中最好的导致3.7倍的荧光蛋白输出和4.4倍的转录水平增加。这些合成终止子提供了优于天然序列的几个优点，包括容易合成的短长度、与天然序列的最小序列同源性以及与常用的较长终止子的性能特征相似或更好的性能特征。此外，合成终止子在酿酒酵母和替代酵母解脂耶氏酵母中都是高度功能性的，表明这些合成设计在不同的酵母物种之间是可转移的。
Terminators play an important role both in completing the transcription process and impacting mRNA half-life. As such, terminators are an important synthetic component considered in applications such as heterologous gene expression and metabolic engineering. Here, we describe a panel of short (35-70 bp) synthetic terminators that can be used for modulating gene expression in yeast. The best of these synthetic terminator resulted in 3.7-fold more fluorescent protein output and 4.4-fold increase in transcript level compared to that with the commonly used CYC1 terminator. These synthetic terminators offer several advantages over native sequences, including an easily synthesized short length, minimal sequence homology to native sequences, and similar or better performance characteristics than those of commonly used longer terminators. Furthermore, the synthetic terminators are highly functional in both Saccharomyces cerevisiae and an alternative yeast, Yarrowia lipolytica, demonstrating that these synthetic designs are transferrable between diverse yeast species.