Luehrsen K R, Pearlman R E, Pata J, Orias E
Department of Biological Sciences, University of California, Santa Barbara, CA 93106.
Curr Genet. 1988 Sep;14(3):225-33. doi: 10.1007/BF00376742.
We have isolated several Tetrahymena thermophila chromosomal DNA fragments which function as autonomously replicating sequences (ARS) in the heterologous Saccharomyces cerevisiae and Schizosaccharomyces pombe selection systems. The Tetrahymena ARS sequences were first isolated in S. cerevisiae and were derived from non-ribosomal micro- and macronuclear DNA. Sequence analysis of the ARS elements identified either perfect or close matches with the 11 bp S. cerevisiae ARS core consensus sequence. Subcloning studies of two Tetrahymena ARS elements defined functional regions ranging in size from 50 to 300 bp. Testing of the ARS elements in S. pombe revealed that most of the T. thermophila inserts confer ARS function in both yeasts, at least in the sense of promoting a high transformation frequency to plasmids which contain them. However, the actual sequences responsible for ARS activity were not always identical in the two yeasts.
我们分离出了几个嗜热四膜虫染色体DNA片段,这些片段在异源酿酒酵母和粟酒裂殖酵母选择系统中作为自主复制序列(ARS)发挥作用。四膜虫ARS序列首先在酿酒酵母中分离得到,来源于非核糖体微核和大核DNA。对ARS元件的序列分析确定了与11 bp酿酒酵母ARS核心共有序列的完美或紧密匹配。对两个四膜虫ARS元件的亚克隆研究确定了大小在50至300 bp之间的功能区域。在粟酒裂殖酵母中对ARS元件的测试表明,大多数嗜热四膜虫插入片段在两种酵母中都赋予ARS功能,至少在促进含有它们的质粒的高转化频率方面是这样。然而,在两种酵母中负责ARS活性的实际序列并不总是相同的。