Assay of biological activity of synthetic yeast alanine transfer RNA (tRNAAlay).
The biological activity of the synthetic tRNAAlay was studied with an extremely sensitive method. tRNAAlay accepted alanine in the presence of rat liver aminoacyl-tRNAAlay-synthetase (this was called the accepting activity). The aminoacylated tRNAAlay was conveniently precipitated by ethanol with good recovery. The efficiency of transferring alanine from the aminoacylated tRNAAlay 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 tRNAAlay 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 tRNAAlay respectively. The efficiency of the incorporation of alanine in the aminoacylated tRNAAlay into the protein was 61.6-63.1%, corresponding to 90.6-91.7% and 97.2-115.8% of that of the natural and the reconstituted natural tRNAAlay respectively.
D000818:Animals; D014176:Protein Biosynthesis; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism / Q000494:pharmacology*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D015003:Yeasts / Q000032:analysis*
D012346:RNA, Transfer, Amino Acyl