Mating response and construction of heterothallic strains of the fission yeast Schizosaccharomyces octosporus
Mating response and construction of heterothallic strains of the fission yeast Schizosaccharomyces octosporus
复制标题
裂殖酵母八孢裂殖酵母的交配反应和异宗合菌株的构建
DOI：
10.1093/femsyr/fox045
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发表时间：
2017
影响因子：
3.2
通讯作者：
Niki Hironori
中科院分区：
文献类型：
--
作者：
Seike Taisuke;Niki Hironori
八孢子裂殖酵母是裂殖酵母属的四个种之一。最近公布的基因组序列数据为比较研究提供了有用的信息。然而，Sz.八孢子虫还没有遗传特征，因为这个物种没有异源菌株。在这里，我们报告了稳定的Sz异源菌株的构建。用于基因杂交的八孢子虫。首先，我们连续观察了同系菌株yFS286的交配过程，并测定了Sz的交配频率。在各种产孢剂上的八孢子菌。结果表明，交配时合子形成率平均为30%，在PMG培养基上合子形成率较高(43.8±44.7%)。八分生裂殖酵母在MEA或YMoA上形成8个孢子的ASCI，但优先产生4个孢子的ASCI。为了获得遗传分析所必需的异硫代菌株，我们分离了表现出异硫代样表型的自发突变株。我们还通过删除沉默区域构建了稳定的异源菌株。因此，我们建立了如下异源菌株TS162ash+和TS150/TS161ash−，并成功地进行了交配。这些遗传工具将用于酵母遗传学，如分子克隆、基因互补测试和四分体(八分体)分析。
The fission yeastSchizosaccharomyces octosporusis one of four species in the genusSchizosaccharomyces. Recently released genome sequence data provide useful information for comparative studies. However,Sz. octosporushas not yet been genetically characterized because there have been no heterothallic strains of this species. Here we report the construction of stable heterothallic strains ofSz. octosporusfor genetic crosses. First, we continuously observed the mating process of a homothallic strain, yFS286, and determined the mating frequency ofSz. octosporuson various sporulation media. It showed, on average, 30% zygote formation on mating, and a higher frequency of zygotes (43.8 ± 4.7%) on PMG medium. Regardless of sporulation, the number of spores within an ascus was variable.Schizosaccharomyces octosporusforms eight-spored asci, but preferentially produced four-spored asci on MEA or YMoA medium. To obtain heterothallic strains essential for genetic analyses, we isolated spontaneous mutants showing heterothallic-like phenotypes. We also constructed stable heterothallic strains by deleting the silentmatregion. As a result, we established the following heterothallic strains, TS162 ash+and TS150/TS161 ash−, which successfully mated with each other. These genetic tools will be useful for yeast genetics such as molecular cloning, gene complementation tests and tetrad (octad) analysis.