Glover D M, Hogness D S
Cell. 1977 Feb;10(2):167-76. doi: 10.1016/0092-8674(77)90212-4.
The sequences corresponding to the 18S and 28S rRNAs have been mapped within a cloned 17 kilobase (kb) fragment formed by Eco R1 cleavage of Drosophila melanogaster rDNA. This fragment, Dm103, represents the longer of two major types of repeating units that are present in the rDNA of this fly, and was cloned as a hybrid plasmid, pDm103, consisting of Dm103 inserted at the Eco R1 site of the pSC101 vector (Glover et al., 1975). Mapping of the 18S and 28S rDNA in Dm103 was accomplished by quantitative determination of the amount of these rDNAs in each member of an ordered set of restriction fragments obtained by Hind III and Eco R1 ccleavage of pDm103. The amounts of 18S and 28S rDNAs were determined by hybridization of the rRNAs to fragments that were purified by cloning, and an unambiguous order of the fragments within pDm103 was established by heteroduplex mapping and from the stoichiometry of the fragment lengths. The resulting map revealed that the 4 kb of 28S rDNA within the long repeating unit represented by Dm103 is divided into two blocks that are separated by 5.4 kb of DNA of unknown function. It is this unusual arrangement of the 28S rDNA that distinguishes the long repeating units (17 kb) from the short units (11.5) kb), whose 4 kb of 28S rDna is confined to a single block, as is shown in the accompanying paper (White and Hogness, 1977). The remainder of the DNA in this long unit appears to be typically arranged, with the 2 kb of 18S rDNA confined to a single block that is separated by about 1 kb from the closest block of 28S rDNA.
与18S和28S核糖体RNA相对应的序列已定位在一个通过用Eco R1切割黑腹果蝇rDNA形成的17千碱基(kb)的克隆片段内。这个片段Dm103代表了这种果蝇rDNA中存在的两种主要重复单元类型中较长的一种,并被克隆为一个杂种质粒pDm103,它由插入到pSC101载体Eco R1位点的Dm103组成(格洛弗等人,1975年)。通过对pDm103经Hind III和Eco R1切割得到的一组有序限制性片段中每个成员所含这些rDNA的量进行定量测定,完成了Dm103中18S和28S rDNA的定位。18S和28S rDNA的量通过将rRNA与通过克隆纯化的片段杂交来测定,并且通过异源双链体定位以及片段长度的化学计量关系确定了pDm103内片段的明确顺序。所得图谱显示,由Dm103代表的长重复单元内的4 kb 28S rDNA被分成两个区域,中间由5.4 kb功能未知的DNA隔开。正是28S rDNA的这种不同寻常的排列方式,将长重复单元(17 kb)与短单元(11.5 kb)区分开来,短单元的4 kb 28S rDNA局限于一个单一区域,如随附论文(怀特和霍格内斯,1977年)所示。这个长单元中其余的DNA似乎排列正常,2 kb的18S rDNA局限于一个单一区域,与最接近的28S rDNA区域相隔约1 kb。