McLachlan A, Hardman N
Biochim Biophys Acta. 1982 Apr 26;697(1):89-100. doi: 10.1016/0167-4781(82)90049-5.
The properties of DNA segments containing foldback elements were studied after their selection from a "random" recombinant library of Physarum polycephalum nuclear DNA sequences, cloned using the plasmid vector pBR322. Hybridisation of in vitro labelled recombinant plasmids to Southern blots of genomic restriction fragments demonstrated that each cloned segment contained repetitive elements located at several hundred sites in the genome. Two of the DNA clones generated hybridisation patterns which suggested that they contain repetitive elements with internal cleavage sites for the restriction endonuclease HaeIII. Cross-hybridisation of all combinations of the cloned sequences showed that most contain different arrangements of repetitive elements derived from different sequence families. The results are consistent with a model proposed previously on the basis of studies on total nuclear DNA, for the organisation of sequences closely associated with foldback elements in the Physarum genome.
从多头绒泡菌核DNA序列的“随机”重组文库中选出含有回文元件的DNA片段,用质粒载体pBR322进行克隆后,对其特性进行了研究。体外标记的重组质粒与基因组限制性片段的Southern印迹杂交表明,每个克隆片段都含有位于基因组数百个位点的重复元件。其中两个DNA克隆产生的杂交模式表明,它们含有对限制性内切酶HaeIII具有内部切割位点的重复元件。克隆序列所有组合的交叉杂交表明,大多数含有来自不同序列家族的重复元件的不同排列。这些结果与先前基于对总核DNA的研究提出的关于多头绒泡菌基因组中与回文元件紧密相关的序列组织的模型一致。