Miao Cui-Cui, Han Lin-Li, Lu Yan-Bing, Feng Hong
Key Laboratory of Molecular Biology and Biotechnology, College of Life Sciences, Sichuan University, 29, Wangjiang Rd, Chengdu 610064, China.
Key Laboratory for Bio-Resources and Eco-Environment of Ministry of Education, Sichuan University, 29 Wangjiang Rd, Chengdu 610064, China.
Microorganisms. 2020 Jul 12;8(7):1030. doi: 10.3390/microorganisms8071030.
is an ideal host for secretion and expression of foreign proteins. The promoter is one of the most important elements to facilitate the high-level production of recombinant protein. To expand the repertoire of strong promoters for biotechnological applications in species, 14 highly transcribed genes based on transcriptome profiling of BA06 were selected and evaluated for their promoter strength in . Consequently, a strong promoter P was obtained, which could drive the genes encoding alkaline protease () and green fluorescent protein (GFP) to express more efficiency by an increase of 3.65-fold and 18.40-fold in comparison with the control promoter (P), respectively. Further, promoter engineering was applied to P, leading to a mutation promoter (P) that could increase GFP expression by 3.67-fold over the wild-type promoter (P). Moreover, the IPTG-inducible expression systems were constructed using the operon based on the strong promoters of P and P, which could work well both in and . In this study, highly efficient expression system for was constructed based on transcriptome data and promoter engineering, which provide not only a new option for recombinant expression in , but also novel genetic tool for .
是外源蛋白分泌和表达的理想宿主。启动子是促进重组蛋白高水平生产的最重要元件之一。为了扩展用于[物种名称]生物技术应用的强启动子库,基于[物种名称]BA06的转录组分析选择了14个高转录基因,并在[具体实验对象]中评估了它们的启动子强度。结果,获得了一个强启动子P,与对照启动子(P)相比,它能驱动编码碱性蛋白酶([蛋白酶名称])和绿色荧光蛋白(GFP)的基因分别以3.65倍和18.40倍的效率更高地表达。此外,对P进行了启动子工程改造,得到了一个突变启动子(P),其能使GFP表达比野生型启动子(P)提高3.67倍。而且,基于P和P的强启动子构建了使用[具体操纵子名称]操纵子的IPTG诱导表达系统,该系统在[具体实验对象1]和[具体实验对象2]中均能良好工作。在本研究中,基于转录组数据和启动子工程构建了[物种名称]的高效表达系统,这不仅为[物种名称]中的重组表达提供了新选择,也为[相关研究领域]提供了新的遗传工具。