Ohtsuki T, Vinayak R, Watanabe Y, Kita K, Kawai G, Watanabe K
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo.
J Biochem. 1996 Dec;120(6):1070-3. doi: 10.1093/oxfordjournals.jbchem.a021522.
RNA samples corresponding to Ascaris suum mitochondrial tRNA(Met) were chemically and automatically synthesized in amounts sufficient for NMR measurement. Conventional and rapid deprotection methods gave tRNA samples with the same amino acid-accepting activity as those prepared by other method; enzymatic synthesis, and enzymatic ligation of chemically synthesized fragments. The synthetic tRNA showed the same 1H-NMR spectrum in the iminoproton region as the ligated tRNA. This rapid and reliable preparation method thus provides biologically active tRNA for NMR measurement, and further, it is applicable for synthesis of other large synthetic RNAs, by combining the site-specific isotopic labeling method.
合成了与猪蛔虫线粒体tRNA(Met)相对应的RNA样本,其化学合成量和自动合成量足以进行核磁共振测量。常规脱保护方法和快速脱保护方法得到的tRNA样本具有与通过其他方法(酶促合成以及化学合成片段的酶促连接)制备的样本相同的氨基酸接受活性。合成的tRNA在亚氨基质子区域的1H-NMR谱与连接的tRNA相同。因此,这种快速可靠的制备方法为核磁共振测量提供了具有生物活性的tRNA,此外,通过结合位点特异性同位素标记方法,它还适用于其他大型合成RNA的合成。