Gottlieb P, LaFauci G, Rudner R
Gene. 1985;33(3):259-68. doi: 10.1016/0378-1119(85)90233-1.
Deletions and additions of rRNA gene sets in Bacillus subtilis were observed by Southern hybridizations using cloned radiolabeled rDNA sequences. Of the ten rRNA gene sets found in B. subtilis 168M or NCTC3610, one was deleted in strains possessing the leuB1, ilvC1, argA2 and pheA1 mutations. Among EcoRI restriction fragments of genomic DNA products, a 2.9-kb 23S rRNA homolog was missing. In HindIII digest, both 5.5- and 5.1-kb hybrid bands were lost with 16S and 23S probes, respectively. Similarly, genomic DNAs digested with SmaI showed the absence of both 2.1- and 2.0-kb fragments that hybridized to 16S and 5S sequences, respectively, in wild-type genomes. In contrast, B. subtilis strain 166 and its derivatives displayed a gain of a 3.3-kb HindIII fragment homologous to 16S rRNA. Transforming the ilvC1 and leuB1 mutations into new genetic backgrounds revealed in some clones the concomitant introduction of the ribosomal defect. Transformations with the slightly heterologous donor DNA from strain W23 yielded some Leu+ and Arg+ transformants with altered hybridization patterns when probed with cloned sequences. We propose that the deletion of the rRNA operon occurred in the ilv-leu gene cluster of the B. subtilis genome as a result of unequal recombination between redundant sequences.
利用克隆的放射性标记rDNA序列进行Southern杂交,观察到枯草芽孢杆菌rRNA基因集的缺失和增加。在枯草芽孢杆菌168M或NCTC3610中发现的10个rRNA基因集中,有一个在具有leuB1、ilvC1、argA2和pheA1突变的菌株中缺失。在基因组DNA产物的EcoRI限制性片段中,一个2.9kb的23S rRNA同源物缺失。在HindIII消化物中,分别用16S和23S探针杂交时,5.5kb和5.1kb的杂交带均消失。同样,用SmaI消化的基因组DNA显示,在野生型基因组中分别与16S和5S序列杂交的2.1kb和2.0kb片段均不存在。相比之下,枯草芽孢杆菌166菌株及其衍生物显示出一个与16S rRNA同源的3.3kb HindIII片段增加。将ilvC1和leuB1突变导入新的遗传背景后,在一些克隆中发现了核糖体缺陷的伴随引入。用来自W23菌株的轻度异源供体DNA进行转化,当用克隆序列进行探测时,产生了一些杂交模式改变的Leu+和Arg+转化体。我们认为,rRNA操纵子的缺失是由于枯草芽孢杆菌基因组中ilv-leu基因簇内冗余序列之间的不等位重组所致。