Douthwaite S, Prince J B, Noller H F
Proc Natl Acad Sci U S A. 1985 Dec;82(24):8330-4. doi: 10.1073/pnas.82.24.8330.
A mutation affording low levels of erythromycin resistance has been obtained by in vitro hydroxylamine mutagenesis of a cloned ribosomal RNA operon from Escherichia coli. The site of the mutational event responsible for antibiotic resistance was localized to the gene region encoding domain II of 23S rRNA by replacement of restriction fragments in the wild-type plasmid by corresponding fragments from the mutant plasmid. DNA sequencing showed that positions 1219-1230 of the 23S rRNA gene are deleted in the mutant. Since all previously characterized rRNA mutations conferring resistance to erythromycin show changes exclusively in domain V, our present findings provide direct evidence for functional interaction between domains II and V of 23S rRNA.
通过对来自大肠杆菌的克隆核糖体RNA操纵子进行体外羟胺诱变，获得了一种具有低水平红霉素抗性的突变。通过用突变体质粒的相应片段替换野生型质粒中的限制片段，将负责抗生素抗性的突变事件位点定位到编码23S rRNA结构域II的基因区域。DNA测序表明，突变体中23S rRNA基因的1219 - 1230位缺失。由于所有先前鉴定的赋予红霉素抗性的rRNA突变仅在结构域V中显示变化，我们目前的发现为23S rRNA结构域II和V之间的功能相互作用提供了直接证据。
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