Dawid I B, Botchan P
Proc Natl Acad Sci U S A. 1977 Oct;74(10):4233-7. doi: 10.1073/pnas.74.10.4233.
Many repeating units of Drosophila melanogaster rDNA contain a DNA sequence within the gene for 28S rRNA that does not code for rRNA. This sequence has been called the ribosomal insertion [Wellauer, P. K. & Dawid, I. B. (1977) Cell 10, 193-212]. We report here that members of the same sequence family occur outside the ribosomal locus. "Non-rDNA insertion DNA" was separated from rDNA by density gradient centrifugation, and sequences homologous to the ribosomal insertion were detected by hybridization with restriction endonuclease fragments derived from a cloned rDNA repeating unit. Pure insertion sequences from cloned rDNA separated from main band DNA and behaved like a component with high G + C content. Non-rDNA components hybridizing to the insertion also separated from main band DNA but less so than pure insertion sequences, suggesting that non-rDNA insertion sequences are linked to DNA of different nucleotide composition. Restriction endonuclease analysis of non-rDNA insertion DNA showed many fragments of different sizes. The patterns obtained were similar in embryonic, larval, pupal, and adult DNA and DNA from cultured cells (Schneider cell line 3). Non-rDNA insertion sequences account for about 0.2% of the genome or about 400 kbases of DNA per haploid complement.
果蝇黑腹种rDNA的许多重复单元在28S rRNA基因内包含一段不编码rRNA的DNA序列。该序列被称为核糖体插入序列[韦劳尔,P.K. & 达维德,I.B.(1977年)《细胞》10卷,193 - 212页]。我们在此报告,同一序列家族的成员出现在核糖体基因座之外。通过密度梯度离心将“非rDNA插入DNA”与rDNA分离,并通过与源自克隆rDNA重复单元的限制性内切酶片段杂交来检测与核糖体插入序列同源的序列。从主带DNA中分离出的克隆rDNA中的纯插入序列,其行为类似于高G + C含量的组分。与插入序列杂交的非rDNA组分也从主带DNA中分离出来,但程度不如纯插入序列,这表明非rDNA插入序列与不同核苷酸组成的DNA相连。对非rDNA插入DNA的限制性内切酶分析显示出许多不同大小的片段。在胚胎、幼虫、蛹和成虫的DNA以及培养细胞(施奈德细胞系3)的DNA中获得的模式相似。非rDNA插入序列约占基因组的0.2%,或每个单倍体基因组约400千碱基的DNA。