Livingston D M
Genetics. 1977 May;86(1):73-84. doi: 10.1093/genetics/86.1.73.
A variety of Saccharomyces strains were examined for the presence of 2micro DNA and, if present, for the pattern of fragments produced by its digestion with site-specific (restriction) endonucleases. Two strains were found that did not contain detectable levels of 2micro DNA, and two strains contained 2micro DNA molecules having only one EcoRI restriction endonuclease recognition site rather than the usual two.-A haploid containing 2micro DNA with one EcoRI restriction site was mated with a haploid containing 2micro DNA with two EcoRI restriction sites and the resulting diploid maintained both types during vegetative growth. Sporulation of the diploid produced four spores, and the clones from these spores contained both types.-A haploid lacking 2micro DNA was mated with a haploid containing 2micro DNA and the resulting diploid contained 2micro DNA. The four clones derived from the haploid spores after sporulation of this diploid all contained 2micro DNA. A rho(-) strain without 2micro DNA was mated to a rho(+) strain with 2micro DNA, and heteroplasmons were selected that had received the nucleus from the strain without 2micro DNA and the mitochondria from the strain with 2micro DNA. Twelve of twenty-four such clones contained 2micro DNA.-I conclude that: (1) the different types of 2micro DNA identified in these strains do not restrict one another, (2) the different types are inherited extrachromosomally, (3) lack of 2micro DNA in two strains is not due to the absence of genes needed for maintenance and (4) the approximately 100 copies of 2micro DNA contained within a single cell are probably clustered within one or a few cytoplasmic organelles.
对多种酿酒酵母菌株进行了检测,以确定是否存在2μm DNA;若存在,则检测其经位点特异性(限制性)内切酶消化后产生的片段模式。发现有两个菌株不含可检测水平的2μm DNA,还有两个菌株所含的2μm DNA分子只有一个EcoRI限制性内切酶识别位点,而非通常的两个。将一个含有一个EcoRI限制性位点的2μm DNA单倍体与一个含有两个EcoRI限制性位点的2μm DNA单倍体进行交配,所得二倍体在营养生长期间保留了两种类型。该二倍体形成的孢子产生了四个孢子,这些孢子的克隆体都包含这两种类型。将一个缺乏2μm DNA的单倍体与一个含有2μm DNA的单倍体进行交配,所得二倍体含有2μm DNA。该二倍体形成孢子后,从单倍体孢子衍生出的四个克隆体都含有2μm DNA。将一个不含2μm DNA的ρ(-)菌株与一个含有2μm DNA的ρ(+)菌株进行交配,并选择了接受了不含2μm DNA菌株的细胞核和含有2μm DNA菌株的线粒体的异质体。在24个这样的克隆体中,有12个含有2μm DNA。我的结论是:(1)在这些菌株中鉴定出的不同类型的2μm DNA不会相互限制;(2)不同类型是通过染色体外遗传的;(3)两个菌株中缺乏2μm DNA并非由于缺乏维持所需的基因;(4)单个细胞内所含的大约100个2μm DNA拷贝可能聚集在一个或几个细胞质细胞器内。