A Robust and Quantitative Reporter System To Evaluate Noncanonical Amino Acid Incorporation in Yeast.

ABSTRACT: Engineering protein translation machinery to incorporate noncanonical amino acids (ncAAs) into proteins has advanced applications ranging from proteomics to single-molecule studies. As applications of ncAAs emerge, efficient ncAA incorporation is crucial to exploiting unique chemistries. We have established a quantitative reporter platform to evaluate ncAA incorporation in response to the TAG (amber) codon in yeast. This yeast display-based reporter utilizes an antibody fragment containing an amber codon at which a ncAA is incorporated when the appropriate orthogonal translation system (OTS) is present. Epitope tags at both termini allow for flow cytometry-based end point readouts of OTS efficiency and fidelity. Using this reporter, we evaluated several factors that influence amber suppres

SUBMITTER: Stieglitz JT 

PROVIDER: S-EPMC6214617 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature