Shostak N G, Kupriianova N S, Serenkova M Ia, Timofeeva M Ia, Baev A A
Mol Biol (Mosk). 1984 Sep-Oct;18(5):1352-62.
The organization of 5S rRNA genes in the loach Misgurnus fossilis L. was studied. 5S rDNA was cloned in Escherichia coli using the HindIII-fragments of loach genome DNA fused into the plasmid pBR322 restricted at the same site. The recombinant clones were tested by colony hybridization. The presence of the 5S rDNA structural sequences in cloned fragments was determined by the Southern procedure of hybridization with 5S [32P]rRNA of the loach and transcription in the oocyte test system. It was found that the size of 5S rDNA repetitive units corresponds to 240-250 bp and 450-460 bp. By the CsCl centrifugation and restriction analysis it was shown that the 5S genes in the loach genome are arranged in clusters (5-30 repeats per cluster), the smaller repeat was found to contain one coding sequence while the larger repeat contains two coding sequences of 5S rDNA.
对泥鳅(Misgurnus fossilis L.)5S rRNA基因的组织进行了研究。利用融合到在同一位置进行酶切的质粒pBR322中的泥鳅基因组DNA的HindIII片段,在大肠杆菌中克隆5S rDNA。通过菌落杂交对重组克隆进行检测。通过与泥鳅的5S [32P]rRNA杂交的Southern杂交程序以及在卵母细胞测试系统中的转录,确定克隆片段中5S rDNA结构序列的存在。发现5S rDNA重复单元的大小对应于240 - 250 bp和450 - 460 bp。通过CsCl密度梯度离心和限制性分析表明,泥鳅基因组中的5S基因成簇排列(每个簇有5 - 30个重复),发现较小的重复包含一个编码序列,而较大的重复包含两个5S rDNA编码序列。