State Key Laboratory of Microbial Technology, Shandong University, Binhai Road 72, Qingdao, Shandong, 266237, People's Republic of China.
Appl Microbiol Biotechnol. 2024 Dec;108(1):14. doi: 10.1007/s00253-023-12835-3. Epub 2024 Jan 3.
The oleaginous yeast Yarrowia lipolytica represents a potential microbial cell factory for the recombinant production of various valuable products. Currently, the commonly used selection markers for transformation in Y. lipolytica are limited, and successive genetic manipulations are often restricted by the number of available selection markers. In our study, we developed a dominant marker, dsdA, which encodes a D-serine deaminase for genetic manipulation in Y. lipolytica. In Y. lipolytica, this marker confers the ability to use D-serine as a nitrogen source. In addition, the selection conditions of several infrequently used dominant markers including bleoR (zeocin resistance), kanMX (G418 resistance), and guaB (mycophenolic acid resistance) were also analyzed. Our results demonstrated that these selection markers can be used for the genetic manipulation of Y. lipolytica and their selection conditions were different for various strains. Ultimately, the selection markers tested here will be useful to expand the genetic toolbox of Y. lipolytica. KEY POINTS: • The dsdA from Escherichia coli was developed as a dominant marker. • The applicability of several resistance markers in Y. lipolytica was determined. • We introduced the Cre/mutant lox system for marker recycling.
油脂酵母解脂耶氏酵母是一种有潜力的微生物细胞工厂，可以用于重组生产各种有价值的产品。目前，用于解脂耶氏酵母转化的常用选择标记物有限，连续的遗传操作经常受到可用选择标记物数量的限制。在我们的研究中，我们开发了一种显性标记物 dsdA，它编码一种 D-丝氨酸脱氨酶，用于解脂耶氏酵母的遗传操作。在解脂耶氏酵母中，该标记赋予了利用 D-丝氨酸作为氮源的能力。此外，还分析了包括 bleoR（博莱霉素抗性）、kanMX（G418 抗性）和 guaB（吗啉代丙氨酸抗性）在内的几个不常用的显性标记物的选择条件。我们的结果表明，这些选择标记物可用于解脂耶氏酵母的遗传操作，其选择条件因不同菌株而异。最终，这里测试的选择标记物将有助于扩展解脂耶氏酵母的遗传工具包。关键点：• 从大肠杆菌中开发出 dsdA 作为显性标记物。• 确定了几种抗性标记物在解脂耶氏酵母中的适用性。• 我们引入了 Cre/突变lox 系统用于标记物回收。
