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Cell-Free Synthesis Of Proteins With Unnatural Amino Acids: Exploring Fitness Landscapes, Engineering Membrane Proteins And Expanding The Genetic Code, Song Min Schinn
Cell-Free Synthesis Of Proteins With Unnatural Amino Acids: Exploring Fitness Landscapes, Engineering Membrane Proteins And Expanding The Genetic Code, Song Min Schinn
Theses and Dissertations
Unnatural amino acids (uAA) expand the structural and functional possibilities of proteins. Numerous previous studies have demonstrated uAA as a powerful tool for protein engineering, but challenges also remain. Three notable such challenges include: (1) the fitness of uAA-incorporated proteins are difficult to predict and time-consuming to screen with conventional methods, (2) uAA incorporation in difficult-to-express proteins (e.g. membrane proteins such as G-protein coupled receptors) remain challenging, and (3) the incorporation of multiple types of uAA are still limited. In response, we pose cell-free protein synthesis (CFPS), a rapid and versatile in vitro expression system, as a platform to explore …
Engineering Cell-Free Protein Synthesis Technology For Codon Reassignment, Biotherapeutics Production Using Just-Add-Water System, And Biosensing Endocrine Disrupting Compounds, Sayed Mohammad Salehi
Engineering Cell-Free Protein Synthesis Technology For Codon Reassignment, Biotherapeutics Production Using Just-Add-Water System, And Biosensing Endocrine Disrupting Compounds, Sayed Mohammad Salehi
Theses and Dissertations
Cell-free protein synthesis is an emerging technology that has many applications. The open nature of this system makes it a compelling technology that can be manipulated to answer many needs that are unavailable in other systems. This dissertation reports on engineering this technology for: 1) sense codon emancipation for incorporation of multiple unnatural amino acids; 2) expressing a hard-to-express anticancer biotherapeutic and introducing a just-add-water system; 3) a biosensing ligand that interacts with nuclear hormone receptors. Emancipating sense codons toward a minimized genetic code is of significant interest to science and engineering. A promising approach to sense codon emancipation is …