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Termination‐Free Prokaryotic Protein Translation by Using Anticodon‐Adjusted E. coli tRNA Ser as Unified Suppressors of the UAA/UGA/UAG Stop Codons. Read‐Through Ribosome Display of Full‐Length DHFR with Translated UTR as a Buried Spacer Arm
Author(s) -
Ogawa Atsushi,
Sando Shinsuke,
Aoyama Yasuhiro
Publication year - 2006
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200500397
Subject(s) - transfer rna , translation (biology) , messenger rna , ribosome , untranslated region , five prime untranslated region , stop codon , protein biosynthesis , polypeptide chain , computational biology , biology , chemistry , rna , amino acid , biochemistry , gene
Don't stop me now . Anticodon‐adjusted tRNA Ser can be used as unified suppressors of the three types of stop codon. In their presence, translation of pseudonatural mRNAs for DHFR is rendered termination‐free and proceeds into the untranslated region (UTR), giving rise to protein–ribosome–mRNA complex with full display of the protein when the translated UTR peptide serving as a spacer arm has a chain‐length of ≥50 amino acids (aa). The mRNA template is recovered from the tag‐selected complex with a 74‐aa UTR in a yield of 2–4 % based on input mRNA.