Replacement of anticodon loop nucleotides to produce functional tRNAs: amber suppressors derived from yeast tRNAPhe.
The method of anticodon loop replacement has been used to make derivatives of yeast tRNAPhe. By constructing tRNAs with a CUA anticodon, complementary to the amber (UAG) terminator, functional amber suppressor tRNAs were produced. The activity of these tRNAs was assayed in a mammalian cell-free protein synthesizing system. The level of suppression reflects the efficiency of codon recognition. tRNAs were constructed with either A, C, U, or G on the 3' side of the CUA anticodon. The tRNAs containing the purines were efficient amber suppressors, whereas those containing pyrimidines were inefficient.
D000926:Anticodon; D001483:Base Sequence; D005815:Genetic Code; D005818:Genetic Engineering; D010649:Phenylalanine; D012343:RNA, Transfer / Q000235:genetics*; D013329:Structure-Activity Relationship; D013489:Suppression, Genetic
D000926:Anticodon; D010649:Phenylalanine; D012343:RNA, Transfer