A new scheme to artificially alter yeast mating-types without autodiploidization
A new scheme to artificially alter yeast mating-types without autodiploidization
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DOI：
10.1016/j.fgb.2020.103442
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发表时间：
2020-11-01
影响因子：
3
通讯作者：
Fukuda, Nobuo
中科院分区：
文献类型：
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作者：
Fukuda, Nobuo
性杂交可以通过从每个亲本细胞遗传基因组信息来驱动酵母的戏剧性进化。然而，不幸的是，出芽酵母的性杂交绝对需要一对菌株，MATa和MAT α交配型，这限制了先前分离和工程菌株的组合可能性。虽然Ho内切酶已被用于人工转化酵母的交配型，但由于DNA消化效率极低，“卡住”突变往往阻碍了交配型转化。为了加速酵母育种技术和菌种工程的发展，需要一种替代的、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。我建立了一种利用MAT基因的合成DNA替代从相反交配型产生MATa和MAT α交配型的方法。我使用了先前构建的episomal载体，该载体抑制现有细胞的交配能力，并产生相反的交配型衍生物，具有抗生素抗性，用于靶细胞分离。我证明了交配型改变的细胞表现出与没有MAT基因替代的单独产生的细胞相同的表型。这种方法可以用较少的努力利用现有资源促进酵母菌株的发育和性杂交。
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 MAT alpha mating-types, is absolutely quired 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 MAT alpha 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.