Nakanishi N, Yamamoto M
Mol Gen Genet. 1984;195(1-2):164-9. doi: 10.1007/BF00332740.
By selecting activities to complement Escherichia coli aro mutations, a gene responsible for the biosynthesis of aromatic amino acids in Schizosaccharomyces pombe (aro3) was cloned into pBR322. Three independent clones named pFNA1, pFNA2 and pFNA3 were obtained. pFNA1 could complement E. coli aroD only, whereas the other two plasmids were able to complement both aroD and aroE. The aro3 locus of S. pombe was found to be a gene cluster which can be subdivided into five complons, A through E (Strauss 1979). Transformation of S. pombe mutants defective in each complon with the pFNA plasmids indicated that pFNA1 carries at least a part of aro3A, the whole of B, C and D, as well as a part of E, whereas pFNA2 and pFNA3 carry a part of aro3C and the entire D and E complons. A physical map of the aro3 locus was constructed by analyzing the structure of these plasmids and their hybridization patterns to restricted genomic DNA. A transcript 4.5 kb long was detected as the sole aro3 mRNA encompassing all five complons. This study, in addition to the work of Strauss (1979), establishes that the arrangement of the aro3 complons in S. pombe in terms of their enzymatic coding activities is identical with that of Neurospora crassa, but different from that proposed for Saccharomyces cerevisiae.
通过选择能互补大肠杆菌aro突变的活性物质,将粟酒裂殖酵母中负责芳香族氨基酸生物合成的一个基因(aro3)克隆到了pBR322中。获得了三个独立的克隆,分别命名为pFNA1、pFNA2和pFNA3。pFNA1只能互补大肠杆菌的aroD,而另外两个质粒能够互补aroD和aroE。发现粟酒裂殖酵母的aro3位点是一个基因簇,可细分为五个互补子,A至E(施特劳斯,1979年)。用pFNA质粒转化每个互补子有缺陷的粟酒裂殖酵母突变体,结果表明pFNA1至少携带aro3A的一部分、整个B、C和D以及E的一部分,而pFNA2和pFNA3携带aro3C的一部分以及整个D和E互补子。通过分析这些质粒的结构及其与限制性基因组DNA的杂交模式,构建了aro3位点的物理图谱。检测到一个4.5 kb长的转录本,作为包含所有五个互补子的唯一aro3 mRNA。除了施特劳斯(1979年)的工作之外,这项研究还证实,粟酒裂殖酵母中aro3互补子在酶编码活性方面的排列与粗糙脉孢菌相同,但与酿酒酵母中提出的排列不同。