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Chapter title |
Directed Evolution of Orthogonal Pyrrolysyl-tRNA Synthetases in Escherichia coli for the Genetic Encoding of Noncanonical Amino Acids
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Chapter number | 5 |
Book title |
Noncanonical Amino Acids
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Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7574-7_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7573-0, 978-1-4939-7574-7
|
Authors |
Moritz J. Schmidt, Daniel Summerer |
Abstract |
The directed evolution of orthogonal aminoacyl-tRNA synthetases (aaRS) for the genetic encoding of noncanonical amino acids (ncAA) has paved the way for the site-specific incorporation of >170 functionally diverse ncAAs into proteins in a large number of organisms [1, 2]. Here, we describe the directed evolution of orthogonal pyrrolysyl-tRNA synthetase (PylRS) mutants with new amino acid selectivities from libraries using a two-step selection protocol based on chloramphenicol and barnase reporter systems. Although this protocol focuses on the evolution of PylRS variants, this procedure can be universally employed to evolve orthogonal aaRS. |
Mendeley readers
The data shown below were compiled from readership statistics for 13 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 13 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Bachelor | 3 | 23% |
Student > Ph. D. Student | 2 | 15% |
Researcher | 2 | 15% |
Other | 1 | 8% |
Unknown | 5 | 38% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 4 | 31% |
Chemistry | 3 | 23% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 8% |
Engineering | 1 | 8% |
Unknown | 4 | 31% |