Isolation of heterothallic haploid and auxotrophic mutants of Schizosaccharomyces japonicus
Isolation of heterothallic haploid and auxotrophic mutants of Schizosaccharomyces japonicus
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DOI：
10.1002/yea.1662
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发表时间：
2009-04-01
期刊：
影响因子：
2.6
通讯作者：
Niki, Hironori
中科院分区：
文献类型：
--
作者：
Furuya, Kanji;Niki, Hironori
裂殖酵母Schizoschiomaceuschivar.粟酒裂殖酵母与粟酒裂殖酵母一起属于裂殖酵母属，粟酒裂殖酵母已经作为模式生物被很好地研究。相比之下，Sz。生物多样性的特征很差，遗传工具尚未开发。我们在这里报告的异宗配合单倍体NIG 2017，NIG 2025和NIG 2028，这是一个Sz的衍生物的分离。同宗配合菌株（NIG 2008）。根据Sz.由布罗德研究所发布，我们发现，Sz。Sz的交配型基因也有直系同源基因。pombe;两个mat-M（-）和两个mat-P（+）基因。正如预期的那样，异宗配合菌株在Sz之一中有缺陷。基因（mat（sj））。我们证实，NIG 2017和NIG 2025菌株仅表达来自mat（sj）-P基因的mRNA，而同宗配合菌株表达mat（sj）-M和mat（sj）-P。尽管NIG 2028菌株表达两种基因产物，但发现mat（sj）-P突变，这可能赋予突变体的异宗配合表型。因此，我们得出结论，这些是稳定的异宗配合菌株。我们将NIG 2017和NIG 2025分别指定为h（+），NIG 2028指定为h（-）。我们还发现了由NIG 2017和NIG 2028衍生物之间的杂交产生的另外的h（-）菌株（NIG 5872和NIG 5873）。除此之外，我们还构建了ura 4（sj）缺失株和ade 6（sj）突变株。利用这些异宗配合菌株和营养缺陷型菌株进行孢子解剖分析，确定了几个位点之间的遗传距离，发现交配型位点和ade 6（sj）位点与着丝粒连锁。版权所有（C）2009约翰威利父子有限公司
The fission yeast Schizosaccharomyces japonicus var. japonicus belong to the genus Schizosaccharomyces, together with Schizosaccharomyces pombe, which has been well studied as a model organism. In contrast, Sz. japonicus is poorly characterized and genetic tools were yet to be developed. We here report the isolation of the heterothallic haploids NIG2017, NIG2025 and NIG2028, which were derivatives of a Sz. japonicus homothallic strain (NIG2008). Based on the genomic sequence of Sz. japonicus, released by the Broad Institute, we found that Sz. japonicus also possesses orthologues of the mating-type genes of Sz. pombe; two mat-M (-) and two mat-P (+) genes. As expected, heterothallic strains were defective in one of the Sz. japonicus mat genes (mat(sj)). We confirmed that NIG2017 and NIG2025 strains only expressed mRNA from the mat(sj)-P genes, while homothallic strains expressed both mat(sj)-M and mat(sj)-P. Although the NIG2028 strain expressed both gene products, mat(sj)-P was found mutated, which may have conferred the heterothallic phenotype of the mutant. Thus, we concluded that these were stable heterothallic strains. We designated NIG2017 and NIG2025 as h(+) and NIG 2028 as h(-), respectively. We also found additional h(-) strains (NIG5872 and NIG5873) that arose from the cross between NIG2017 and NIG2028 derivatives. In addition to that, we have constructed a ura4(sj)-deleted strain and an ade6(sj)-mutated strain. We used these heterothallic strains and the auxotroph strains to perform spore dissection analysis to determine the genetic distances between several loci, and found that the mating type loci and ade6(sj) locus were linked to centromeres. Copyright (C) 2009 John Wiley & Sons, Ltd.