Codon recognition by artificial tRNA molecules with modified nucleosides in the anticodon

Proteins with unnatural amino acids at specific positions can be produced through cell-free protein synthesis. The synthesis of such molecules can, in principle, be facilitated by improving the codon reading efficiency of the tRNA that inserts the unnatural amino acid. In the present study, we prepared tRNA molecules with 2'-O-methyl nucleosides at the second and third positions of the anticodon and measured their codon-reading efficiencies. The results indicated, contrary to our expectation, that the modification damaged the decoding function completely.