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| SYNTHESIS OF THE 5'-HALF MOLECULE OF YEAST ALANINE tRNA | |
| 作者姓名： | 王德宝 吴仁龙 郑可沁 于大文 汤锦炎 祁国荣 陈常庆 朱莹书 谢恒月 王应魁 聂慧玲 陈海宝 黄建华 陈德化 庄建华 王贵海 陈慎 |
| 作者单位： | Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Cell Biology，Academia Sinica，Shanghai Institute of Cell Biology，Academia Sinica，Shanghai Institute of Cell Biology，Academia Sinica，Shanghai Institute of Cell Biology，Academia Sinica，Shanghai Institute of Organic Chemistry，Academia Sinica，Shanghai Institute of Organic Chemistry，Academia Sinica，Shanghai Institute of Organic Chemistry，Academia Sinica，Shanghai Institute of Organic Chemistry，Academia Sinica，Tnstitute of Biophysics，Academia Sinica，Beijing，Tnstitute of Biophysics，Academia Sinica，Beijing |
| 摘 要： | In this paper, we report the synthesis of the 5'-half molecule of yeast alanine tRNA (tRNA_y~(Ala)) by ligating three oligonucleotide fragments corresponding to the nucleotide sequences 1-13, 14-22 and 23-35 respectively under the catalysis of T_4 RNA ligase (Fig. 1). Because of the high purity of the oligonuclcotide fragments and the excellent quality of T_4RNA ligase and polynucleotide kinase we prepared, the isolation steps were simplified and the overall yields were much higher. The ligating yield of the docosamer (Ⅳ) was 75%, that of the pentatriacontamer (Ⅴ), 90%, and the isolated yield of the final product was 21% calculated on the basis of the tridecamer (Ⅲ) used in the. first reaction. Under the action of T_4 RNA ligase the synthetic 5'-half molecule was joined with the natural 3'-half molecule forming a semi-synthetic tRNA_y~(Ala), which possessed the biological activities of both accepting (~3H)-alanine and incorpprating it into proteins. The correctness of the structure of the synthetic 5 |
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