Chakraburtty K
Nucleic Acids Res. 1980 Oct 10;8(19):4459-72. doi: 10.1093/nar/8.19.4459.
Escherichia coli tRNAArg was digested with ribonuclease T1 under restrictive conditions in order to dissect a minimum number of diester bonds. The number of diester bonds cleaved and their locations were determined by phosphorylation of the newly formed 5' hydroxyl groups with [32P] ATP and polynucleotide kinase. There was complete loss of aminoacylation of tRNAARg when two diester bonds were cleaved at the anticodon. However, this material retained the specific properties of synthetase recognition. Two fragments were derived by further digestion of this tRNA. One 19 nucleotide-long fragment derived from the 3' end of tRNAArg and another 18 nucleotide-long fragment derived from the 5' end of the molecule were required to maintain the properties of the specific recognition by the arginyl tRNA synthetase in the absence of the rest of the structure including the anticodon.
在严格条件下用核糖核酸酶T1消化大肠杆菌tRNAArg，以便剖析最少数量的磷酸二酯键。通过用[32P]ATP和多核苷酸激酶对新形成的5'羟基进行磷酸化来确定被切割的磷酸二酯键的数量及其位置。当在反密码子处切割两个磷酸二酯键时，tRNAARg的氨酰化完全丧失。然而，这种物质保留了合成酶识别的特异性。通过对这种tRNA的进一步消化得到两个片段。在没有包括反密码子在内的其余结构的情况下，为了维持精氨酰tRNA合成酶特异性识别的特性，需要一个来自tRNAArg 3'端的19个核苷酸长的片段和另一个来自分子5'端的18个核苷酸长的片段。
