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| SYNTHESIS OF THE 3'-HALF MOLECULE OF YEAST ALANINE tRNA | |
| 作者姓名： | 王德宝 裘慕绥 梁镇和 郑可沁 吴仁龙 汪成尧 刘新垣 郑宏大 包永德 朱心良 郭礼和 陈雅娣 黄敬坚 周凤仪 陈海宝 徐保中 张其玖 华陵 胡美浩 |
| 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 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 Organio Chemistry，Academia Sinica，Shanghai Institute of Organio Chemistry，Academia Sinica，Shanghai Institute of Organio Chemistry，Academia Sinica，Shanghai Institute of Organio Chemistry，Academia Sinica，Institute of Biophysics，Academia Sinica，Beijing，Institute of Biophysics，Academia Sinica，Beijing，Department of Biology，Peking University |
| 摘 要： | This paper deals with the synthesis of the 3'-half molecule of yeast alanine transfer RNA (tRNA_y~(Ala)) by ligation with T_4 RNA ligase of three component oligonucleotide fragments corresponding to nucleotides 36-45(Ⅰ), 46-57(Ⅱ) and 58-76(Ⅲ) in succession extending from the 3'-end to the 5'-end. First, in a ratio of acceptor to donor at 1.5 to 1, we adopted a method of three successive reactions, namely, the 5'-phosphorylation of the nonadecamer (Ⅲ), ligation with the dodecamer(Ⅱ) and the 5'-phosphorylation of the ligation product formed; with one isolation step and obtained the 5'-phosphorylated 31mer(46-76) (Ⅳ) in an overall yield of 70%. Then the 31met(Ⅵ) as a donor was ligated with 3 times of decamer(Ⅰ) to form the 41met(36-76) (Ⅴ), the 3'-half molecule of tRNA_y~(Ala)). The yield was 67%. After 5'-phosphorylation, (Ⅴ) was ligated with the natural 5'-half molecule to form the semi-synthetic tRNA_y~(Ala)), which was biologically active, i. e. accepting and transferring (~3H)-alanine into p |
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