Nagasaki A, Sutoh K, Adachi H, Sutoh K
Department of Life Science, Graduate School of Arts and Sciences, University of Tokyo, Japan.
Biochem Biophys Res Commun. 1998 Mar 17;244(2):505-13. doi: 10.1006/bbrc.1998.8230.
Using a method of random insertional mutagenesis called REMI (restriction enzyme-mediated integration), we isolated two mutant strains of Dictyostelium discoideum with a defect in cAMP-dependent cell aggregation. On bacterial lawns, both of the cells formed large and smooth plaques. When starved in a non-nutrient medium, they became elongated and extended pseudopods very frequently like starved wild type cells. However, they never formed streams toward an aggregation center. Genomic DNA fragments flanking the sites of insertion of the REMI tag were rescued from the mutant cells. The fragments contained one common open reading frame encoding a protein of 1148 amino acid residues. The protein's sequence is homologous to those of two hypothetical proteins of S. cerevisiae and S. pombe.
我们使用一种名为REMI（限制性内切酶介导的整合）的随机插入诱变方法，分离出了两株盘基网柄菌的突变菌株，它们在依赖cAMP的细胞聚集方面存在缺陷。在细菌平板上，这两种细胞都形成了大而光滑的菌斑。当在无营养培养基中饥饿时，它们会变得细长，并像饥饿的野生型细胞一样频繁地伸出伪足。然而，它们从未形成朝向聚集中心的细胞流。从突变细胞中拯救出了REMI标签插入位点两侧的基因组DNA片段。这些片段包含一个共同的开放阅读框，编码一个由1148个氨基酸残基组成的蛋白质。该蛋白质的序列与酿酒酵母和粟酒裂殖酵母的两种假定蛋白质的序列同源。
