Bozsik Aniko, Szilagyi Z, Benko Z, Sipiczki M
Department of Genetics, University of Debrecen, Debrecen, Hungary.
Yeast. 2002 Apr;19(6):485-98. doi: 10.1002/yea.853.
The dimorphic fission yeast Schizosaccharomyces japonicus has proved to be an excellent experimental model for the investigation of the eukaryotic cell. Here we show that it has a haplontic life cycle, in which the diploid phase is confined to the zygote. To make it amenable to genetic and molecular analysis, we generated genetic markers and cloned a genomic sequence which acts as ars when integrated into a plasmid. Diploids suitable for testing complementation and recombination between markers can be formed by protoplast fusion. The complementation tests and the recombination frequencies determined in octads of spores identified 28 non-allelic groups (genes) of mutations of the auxotrophic and mycelium-negative mutants. Two groups of linked markers were also identified. The cloned fragment, which expresses ars activity, encodes a putative amino acid sequence highly similar to a conserved domain of proteins Cut1 (Schizosaccharomyces pombe), BimB (Aspergillus nidulans) and Esp1 (Saccharomyces cerevisiae).
二型裂殖酵母日本裂殖酵母已被证明是研究真核细胞的优秀实验模型。在此我们表明它具有单倍体生命周期，其中二倍体阶段局限于合子。为使其适用于遗传和分子分析，我们生成了遗传标记并克隆了一个基因组序列，该序列整合到质粒中时可作为自主复制序列（ARS）发挥作用。通过原生质体融合可形成适合测试标记之间互补和重组的二倍体。在孢子八分子中确定的互补测试和重组频率鉴定出了28个非等位基因群（基因），这些基因来自营养缺陷型和菌丝体阴性突变体的突变。还鉴定出了两组连锁标记。表达ARS活性的克隆片段编码一个推测的氨基酸序列，该序列与蛋白质Cut1（粟酒裂殖酵母）、BimB（构巢曲霉）和Esp1（酿酒酵母）的保守结构域高度相似。
