Wang T T, Choi Y J, Lee B H
Department of Food Science and Agricultural Chemistry, McGill University, Ste. Anne de Bellevue, Quebec, Canada.
Crit Rev Biotechnol. 2001;21(3):177-218. doi: 10.1080/20013891081719.
This review describes the transformation systems including vectors, replicons, genetic markers, transformation methods, vector stability, and copy numbers of 13 genera and 31 species of non-Saccharomyces yeasts. Schizosaccharomyces pombe was the first non-Saccharomyces yeast studied for transformation and genetics. The replicons of non-Saccharomyces yeast vectors are from native plasmids, chromosomal DNA, and mitochondrial DNA of Saccharomyces cerevisiae, non-Saccharomyces yeasts, protozoan, plant, and animal. Vectors such as YAC, YCp, YEp, YIp, and YRp were developed for non-Saccharomyces yeasts. Forty-two types of genes from bacteria, yeasts, fungi, and plant were used as genetic markers that could be classified into biosynthetic, dominant, and colored groups to construct non-Saccharomyces yeasts vectors. The LEU2 gene and G418 resistance gene are the two most popular markers used in the yeast transformation. All known transformation methods such as spheroplast-mediating method, alkaline ion treatment method, electroporation, trans-kingdom conjugation, and biolistics have been developed successfully for non-Saccharomyces yeasts, among which the first three are most widely used. The highest copy number detected from non-Saccharomyces yeasts is 60 copies in Kluyveromyces lactis. No general rule is known to illustrate the transformation efficiency, vector stability, and copy number, although factors such as vector composition, host strain, transformation method, and selective pressure might influence them.
本综述描述了包括载体、复制子、遗传标记、转化方法、载体稳定性以及13个属和31种非酿酒酵母的拷贝数在内的转化系统。粟酒裂殖酵母是首个被研究用于转化和遗传学研究的非酿酒酵母。非酿酒酵母载体的复制子来自酿酒酵母、非酿酒酵母、原生动物、植物和动物的天然质粒、染色体DNA和线粒体DNA。已开发出如YAC、YCp、YEp、YIp和YRp等载体用于非酿酒酵母。来自细菌、酵母、真菌和植物的42种基因被用作遗传标记，可分为生物合成、显性和显色组以构建非酿酒酵母载体。LEU2基因和G418抗性基因是酵母转化中最常用的两种标记。所有已知的转化方法，如原生质球介导法、碱性离子处理法、电穿孔法、跨界接合和生物弹道法，均已成功开发用于非酿酒酵母，其中前三种方法应用最为广泛。在乳酸克鲁维酵母中检测到的非酿酒酵母的最高拷贝数为60个拷贝。尽管载体组成、宿主菌株、转化方法和选择压力等因素可能会影响转化效率、载体稳定性和拷贝数，但目前尚无普遍规律可循。
