van den Berg M A, Steensma H Y
Department of Microbiology and Enzymology, Kluyver Laboratory for Biotechnology, Delft University of Technology, The Netherlands.
Yeast. 1997 May;13(6):551-9. doi: 10.1002/(SICI)1097-0061(199705)13:6<551::AID-YEA113>3.0.CO;2-0.
We employed two genes in constructing yeast expression cassettes for dominant, selectable markers. The Saccharomyces cerevisiae gene SFA1, encoding formaldehyde dehydrogenase, was placed under the control of the GPD1 promoter and CYC1 terminator. The Moraxella sp. strain B gene dehH1, encoding fluoroacetate dehalogenase, was placed under the control of both the GPD1 and CYC1 promoters. With these constructs it was possible to select directly for yeast strains resistant to either formaldehyde or fluoroacetate. Both selective agents are completely metabolized and inexpensive, making them very useful in the pursuit of yeast gene functions and for industrial applications. An additional advantage of the formaldehyde dehydrogenase marker is that it is an S. cerevisiae gene, thus allowing 'all yeast' constructs.
我们使用了两个基因来构建用于显性选择标记的酵母表达盒。酿酒酵母基因SFA1编码甲醛脱氢酶，其置于GPD1启动子和CYC1终止子的控制之下。莫拉克斯氏菌属菌株B的基因dehH1编码氟乙酸脱卤酶，其置于GPD1和CYC1启动子的控制之下。利用这些构建体，可以直接筛选出对甲醛或氟乙酸具有抗性的酵母菌株。这两种选择剂都能被完全代谢且价格低廉，使其在研究酵母基因功能和工业应用方面非常有用。甲醛脱氢酶标记的另一个优点是它是酿酒酵母基因，因此可以构建“全酵母”构建体。
