Sexual hybridization can drive dramatic yeast evolution through the inheritance of genomic information from each parental cell. Unfortunately, however, a pair of strains, MATa and MATα mating-types, is absolutely required for sexual hybridization of budding yeasts, which restricts the combining possibilities of previously isolated and engineered strains. While the Ho endonuclease has been used to artificially convert yeast mating-types, “stuck” mutations are often hindrances to mating-type conversion due to extremely low efficiency of DNA digestion. An alternative and powerful approach to generate mating strains with specific properties and desired mating-type from existing strains is needed, to accelerate progress in yeast breeding technology and strain engineering. I established an approach for generating MATa and MATα mating-types from those of the opposite mating-type using synthetic DNA substitution of the MAT gene. I used previously constructed episomal vectors that suppress the mating ability of existing cells and produce opposite mating-type derivatives with antibiotic resistance for target cell isolation. I demonstrated that the mating-type-altered cells exhibited the same phenotype as those separately generated without MAT gene substitution. This approach can facilitate yeast-strain development and sexual hybridization using available resources with less efforts.

有性杂交可以通过从每个亲本细胞继承基因组信息来推动酵母的戏剧性进化。然而不幸的是，一对菌株MAT a和MATα 交配型是芽殖酵母的有性杂交绝对需要的，这限制了先前分离和工程菌株的结合可能性。虽然 Ho 核酸内切酶已被用于人工转换酵母交配型，但由于 DNA 消化效率极低，“卡住”突变通常会阻碍交配型转换。需要另一种强大的方法来从现有菌株中产生具有特定特性和所需交配类型的交配菌株，以加速酵母育种技术和菌株工程的进步。我建立用于产生方法MAT一个和MAT使用的合成的DNA替代从那些相反交配型的α交配类型MAT基因。我使用先前构建的附加型载体抑制现有细胞的交配能力并产生具有抗生素抗性的相反交配型衍生物用于靶细胞分离。我证明了交配类型改变的细胞表现出与那些单独产生的没有MAT基因替换的细胞相同的表型。这种方法可以使用可用资源以较少的努力促进酵母菌株的发育和有性杂交。