Furuya Kanji, Niki Hironori
Radiation Biology Center, Kyoto University, Yosida-Konoe-Cho, Sakyo-Ku, Kyoto 606-8501, Japan.
Microbial Genetics Laboratory, Genetic Strains Research Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan
Cold Spring Harb Protoc. 2017 Dec 1;2017(12):pdb.prot091843. doi: 10.1101/pdb.prot091843.
Haploid yeast cells mate to form heterozygotes and subsequently undergo meiosis to form spores. This process can be used to produce gene combinations and variants that are useful for genetic analysis. For example, these spores can be used to generate double mutants or to measure genetic distances in a mutational analysis. Here, we describe mating and spore dissection procedures for cells. Although the overall procedures resemble those used in , some differences exist, including the use of EMM2 medium without nitrogen (EMM-N) for mating and the shorter incubation time of 16-20 h for cells. Furthermore, the zygotes produce eight spores and thus require an "octad" analysis.
单倍体酵母细胞通过交配形成杂合子，随后进行减数分裂形成孢子。这个过程可用于产生对遗传分析有用的基因组合和变体。例如，这些孢子可用于产生双突变体或在突变分析中测量遗传距离。在这里，我们描述了细胞的交配和孢子解剖程序。尽管总体程序与中使用的程序相似，但存在一些差异，包括使用不含氮的EMM2培养基（EMM-N）进行交配以及细胞16 - 20小时的较短孵育时间。此外，合子产生八个孢子，因此需要进行“八分子”分析。
