Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
Methods Mol Biol. 2021;2196:1-13. doi: 10.1007/978-1-0716-0868-5_1.
The use of the budding yeast Saccharomyces cerevisiae as a model genetic organism has been facilitated by the availability of a wide range of yeast shuttle vectors, plasmids that can be propagated in Escherichia coli and also in yeast, where they are stably maintained at low- or high-copy number, depending on the plasmid system. Here we provide an introduction to the low-copy (ARS/CEN) and multi-copy (2-μm-based) plasmids, the marker genes commonly used for plasmid selection in yeast, methods for transforming yeast and monitoring plasmid inheritance, and tips for working with yeast transformants.
使用 budding 酵母 Saccharomyces cerevisiae 作为模式遗传生物，得益于广泛的酵母穿梭载体的可用性，这些载体可以在 Escherichia coli 和酵母中增殖，并且可以在酵母中稳定地维持低拷贝数或高拷贝数，具体取决于质粒系统。在这里，我们将介绍低拷贝数（ARS/CEN）和多拷贝数（2-μm 为基础）质粒、酵母中质粒选择常用的标记基因、酵母转化方法和质粒遗传监测方法，以及处理酵母转化体的技巧。
