Organization of Advanced Science and Technology, Kobe University, Kobe, Japan.
FEMS Yeast Res. 2014 May;14(3):399-411. doi: 10.1111/1567-1364.12138. Epub 2014 Feb 10.
Yeast has the potential to be used in bulk-scale fermentative production of fuels and chemicals due to its tolerance for low pH and robustness for autolysis. However, expression of multiple external genes in one host yeast strain is considerably labor-intensive due to the lack of polycistronic transcription. To promote the metabolic engineering of yeast, we generated systematic and convenient genetic engineering tools to express multiple genes in Saccharomyces cerevisiae. We constructed a series of multi-copy and integration vector sets for concurrently expressing two or three genes in S. cerevisiae by embedding three classical promoters. The comparative expression capabilities of the constructed vectors were monitored with green fluorescent protein, and the concurrent expression of genes was monitored with three different fluorescent proteins. Our multiple gene expression tool will be helpful to the advanced construction of genetically engineered yeast strains in a variety of research fields other than metabolic engineering.
酵母由于其耐受低 pH 值和自溶的稳健性，有可能用于大规模发酵生产燃料和化学品。然而，由于缺乏多顺反子转录，在一个宿主酵母菌株中表达多个外源基因是相当费力的。为了促进酵母的代谢工程，我们开发了系统和方便的遗传工程工具，以在酿酒酵母中表达多个基因。我们构建了一系列多拷贝和整合载体，通过嵌入三个经典启动子，在酿酒酵母中同时表达两个或三个基因。通过绿色荧光蛋白监测构建载体的比较表达能力，并通过三种不同的荧光蛋白监测基因的同时表达。我们的多基因表达工具将有助于除代谢工程以外的各种研究领域中遗传工程酵母菌株的高级构建。
