ASSAY OF BIOLOGICAL ACTIVITY OF SYNTHETIC YEAST ALANINE TRANSFER RNA (tRNA_y~(Ala))

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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 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 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 results in the assay of 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 6 1.6-63.1%, corresp
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