Sakaguchi J, Yamamoto M
Proc Natl Acad Sci U S A. 1982 Dec;79(24):7819-23. doi: 10.1073/pnas.79.24.7819.
DNA segments cloned from Schizosaccharomyces pombe by the ability to complement Escherichia coli pyrB mutations are shown to complement a ural mutation in S. pombe, thereby demonstrating that ural is the structural gene for aspartate transcarbamylase of S. pombe. Further, such segments combined with parts or all of pBR322 are shown to be capable of autonomous propagation in S. pombe. This suggests the existence of an autonomously replicating sequence (ars) in the vicinity of ural. Unlike the TRP1 segment cloned from Saccharomyces cerevisiae [Struhl, K., Stinchcomb, D. T., Scherer, S. & Davis, R. W. (1979) Proc. Natl. Acad. Sci. USA 76, 1035-1039], plasmids carrying the ural locus do not multiply as monomers but assume a polymeric form as large as a decamer to an icosamer in the yeast. Monomers are tandemly arranged in the polymer. Inversion of an inserted fragment or insertion of another segment into a competent plasmid greatly decreases the efficiency of such transformation, implying a role of the tertiary structure of the plasmids in the establishment of transformation of this kind.
通过互补大肠杆菌pyrB突变的能力从粟酒裂殖酵母中克隆的DNA片段,被证明可互补粟酒裂殖酵母中的ura1突变,从而证明ura1是粟酒裂殖酵母天冬氨酸转氨甲酰酶的结构基因。此外,这类片段与部分或全部pBR322组合后,被证明能够在粟酒裂殖酵母中自主增殖。这表明在ura1附近存在一个自主复制序列(ars)。与从酿酒酵母中克隆的TRP1片段不同[斯特鲁尔,K.,斯廷科姆,D. T.,谢勒,S. & 戴维斯,R. W.(1979年)美国国家科学院院刊76,1035 - 1039],携带ura1位点的质粒在酵母中不是以单体形式增殖,而是呈现出多聚体形式,大小从十聚体到二十聚体不等。单体在多聚体中呈串联排列。将插入片段进行反向操作或在合适的质粒中插入另一个片段会大大降低这种转化的效率,这意味着质粒的三级结构在这类转化的建立中发挥作用。