| ASSAY OF BIOLOGICAL ACTIVITY OF SYNTHETIC YEAST ALANINE TRANSFER RNA (tRNA_y~(Ala)) |
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| 作者姓名： | 申庆祥 余允华 张孝勇 郭礼和 匡达人 包俊茹 谌章群 |
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| 作者单位： | 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 Cell Biology，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica，Shanghai Institute of Biochemistry，Academia Sinica |
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| 摘 要： | The biological activity of the synthetic tRNA_y~(Ala) was studied with an extremely sensitive method, tRNA_y~(Ala). accepted alanine in the presence of rat liver aminoacyl-tRNA_y~(Ala)-synthetase (this was called the accepting actvity). The aminoacylated tRNA_y~(Ala) was conveniently precipitated by ethanol with good recovery. The efficiency of transferring alanine from the aminoacylated tRNA_y~(Ala) into the protein was determined in in vitro rabbit reticulocyte lysate cell-free protein-synthesizing system (this was called the incorporation activity). Both accepting and incorporation activities could be determined in one assay with only 5-7 pmoles of tRNA_y~(Ala) either in ligation mixture or in purified form.Our results show that the accepting activities of the synthetic products were 51.6-65.6% and 91.3-106.0% of that of natural and reconstituted natural tRNA_y~(Ala) respectively. The efficiency of the incorporation of alanine in the aminoacylated tRNA_y~(Ala) into the protein was 61.6-63.1%, corresp
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