Compositions and Methods Of Use Thereof For Making Polypeptides With Many Instances Of Nonstandard Amino Acids

摘要

Compositions, systems, and methods for preparation of polypeptides having multiple iterations of non-standard amino acids are provided. The compositions and method can be used to produce recombinant proteins at a greater yield than the same or similar polypeptides made using conventional compositions, systems, and methods. Accordingly, in some embodiments, the polypeptides are ones that could not be made using conventional methods and reagents, or could not be made a sufficient yield or purity to serve a practical purpose using conventional methods and reagents. Polypeptides made using the disclosed compositions, systems, and methods are also provided.

主权项

1. A method of making aminoacyl tRNA synthetases (AARS) variants comprising
carrying out one or more rounds of mutagenesis on host organism cells comprising an orthogonal translation system comprising a precursor heterologous non-standard amino acid anaminoacyl tRNA synthetase (AARS) integrated into the host's genome and a cognate tRNA to form a library of variant AARS, carrying out one or more rounds of selection on the library of variants to identify host cells comprising variant AARS with improved or altered specificity and/or activity for the non-standard amino and/or the cognate tRNA.