Ando H, Kitabatake M, Inokuchi H
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
Genes Genet Syst. 1996 Feb;71(1):47-50. doi: 10.1266/ggs.71.47.
From Escherichia coli cells with a deletion in the ssrA gene that encodes 10Sa RNA after treatment with a mutagen, we isolated two temperature-sensitive mutants, which we designated TS15 and TS101. The temperature-sensitive (ts) phenotype of the mutants could be overcome by introduction of the wild-type ssrA gene but not by the mutants of ssrA. By a complementation test using Kohara's mini-set of clones and by subcloning of a fragment from the phage clone 246, we found that both mutations were in the prs gene that encodes phosphoribosyl pyrophosphate synthetase. Sequencing of the mutant prs gene of TS101 showed that residues 215, cysteine, in the encoded protein had been changed to tyrosine. That such a mutant exists suggests that 10Sa RNA associate with the prs gene product in a functional way.
在用诱变剂处理后，从编码10Sa RNA的ssrA基因缺失的大肠杆菌细胞中，我们分离出了两个温度敏感突变体，分别命名为TS15和TS101。这些突变体的温度敏感（ts）表型可以通过导入野生型ssrA基因来克服，但不能通过ssrA的突变体来克服。通过使用小原的小克隆集进行互补试验以及对噬菌体克隆246的一个片段进行亚克隆，我们发现这两个突变都位于编码磷酸核糖焦磷酸合成酶的prs基因中。对TS101的突变prs基因进行测序表明，编码蛋白中的第215位残基半胱氨酸已变为酪氨酸。这样一个突变体的存在表明10Sa RNA以一种功能性方式与prs基因产物相关联。
