Meeker R, Tewari K K
Biochemistry. 1980 Dec 23;19(26):5973-81. doi: 10.1021/bi00567a005.
The saturation hybridization between pea ctDNA and 125I-labeled pea ct-tRNAs has shown that 1.2% of the peak ctDNA codes for tRNA genes. The observed level of hybridization has been found to result from specific base pairings between ctDNA and ct-RNA as shown by competition hybridization experiments and thermal stability studies on DNA-tRNA hybrids. The level of hybridization obtained in this study amounts to the presence of approximately 40 tRNA genes in pea ctDNA. The tRNAs from the cytoplasm of the pea leaves, Escherichia coli, yeast, and calf thymus did not compete with the pea ct-tRNAs for the common base sequences in pea ctDNA. The presence of 17 aminoacyl-tRNA synthetases and their corresponding tRNAs was demonstrated in chloroplast. The acylation of ct-tRNAs proceeds with the same rate whether the partially purified tRNA synthetases from chloroplasts of E. coli are used. The aminoacylation of the three amino acids glutamic acid, glutamine, and cysteine proceeded very slowly in chloroplasts. The individually labeled aminoacyl-tRNAs hybridized with pea ctDNA. The hybridization follows true saturation rates, and the melting profiles of aminoacyl-tRNA-ctDNA indicate the formation of specific base pairs between the ctDNA and tRNA. Seventeen aminoacyl-tRNA genes have been identified in the pea ctDNA.
豌豆叶绿体DNA(ctDNA)与125I标记的豌豆叶绿体转运RNA(ct - tRNAs)之间的饱和杂交实验表明,1.2%的峰值ctDNA编码tRNA基因。通过竞争杂交实验以及对DNA - tRNA杂交体的热稳定性研究发现,观察到的杂交水平是由ctDNA与ct - RNA之间的特异性碱基配对导致的。本研究中获得的杂交水平相当于豌豆ctDNA中大约存在40个tRNA基因。豌豆叶片细胞质、大肠杆菌、酵母和小牛胸腺的tRNA不会与豌豆ct - tRNAs竞争豌豆ctDNA中的共同碱基序列。在叶绿体中证实了17种氨酰 - tRNA合成酶及其相应tRNA的存在。无论使用从大肠杆菌叶绿体中部分纯化的tRNA合成酶,ct - tRNAs的酰化反应速率都相同。在叶绿体中,谷氨酸、谷氨酰胺和半胱氨酸这三种氨基酸的氨酰化反应进行得非常缓慢。单独标记的氨酰 - tRNAs与豌豆ctDNA杂交。杂交遵循真正的饱和速率,氨酰 - tRNA - ctDNA的解链曲线表明ctDNA与tRNA之间形成了特异性碱基对。在豌豆ctDNA中已鉴定出17个氨酰 - tRNA基因。