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通过在 Schizosaccharomyces japonicus 中等位基因间互补修复 ade6 构建二倍体合子。

Construction of diploid zygotes by interallelic complementation of ade6 in Schizosaccharomyces japonicus.

机构信息

Microbial Genetics Laboratory and Department of Genetics, Genetic Strains Research Centre, National Institute of Genetics, Shizuoka, Japan.

出版信息

Yeast. 2011 Oct;28(10):747-54. doi: 10.1002/yea.1898. Epub 2011 Sep 8.

Abstract

The construction of diploid cells eases genetic analysis in haploid genetic systems because diploid cells allow for the characterization of essential genes. Here, we report the construction of diploid cells using ade6 point mutants that suppress each other via interallelic complementation in the fission yeast Schizosaccharomyces japonicus var japonicus (Sz. japonicus). We constructed an ade6-domK mutant in addition to the previously described ade6-domE. Phenotypes of both mutants exhibited adenine auxotrophy and red colonies. The mutations complemented the phenotypes in a mutually dependent manner. Diploid zygotes, in which the two mutations were introduced simultaneously into the same cells, were isolated by selecting for adenine independence. Such diploid cells are apparently larger in size than haploid cells, yet have a similar nuclear/cytoplasmic ratio, and thus the nuclear size control that has been reported in Sz. pombe is also present in Sz. japonicus.

摘要

二倍体细胞的构建简化了单倍体遗传系统中的遗传分析,因为二倍体细胞允许对必需基因进行特征描述。在这里,我们报告了使用 ade6 点突变体在裂殖酵母 Schizosaccharomyces japonicus var japonicus(Sz. japonicus)中通过等位基因间互补来构建二倍体细胞。除了之前描述的 ade6-domE 之外,我们还构建了 ade6-domK 突变体。两个突变体的表型均表现为腺嘌呤营养缺陷型和红色菌落。这些突变以相互依赖的方式互补表型。通过选择腺嘌呤独立性,从同一细胞中同时引入两种突变的二倍体合子被分离出来。这样的二倍体细胞明显比单倍体细胞大,但具有相似的核/细胞质比,因此在 Sz. pombe 中报道的核大小控制也存在于 Sz. japonicus 中。

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