A General Strategy for Site-specific Incorporation of Unnatural Amino Acids Into Proteins

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ISBN 13 :
Total Pages : 676 pages
Book Rating : 4.:/5 (33 download)

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Book Synopsis A General Strategy for Site-specific Incorporation of Unnatural Amino Acids Into Proteins by : Spencer Jay Anthony-Cahill

Download or read book A General Strategy for Site-specific Incorporation of Unnatural Amino Acids Into Proteins written by Spencer Jay Anthony-Cahill and published by . This book was released on 1990 with total page 676 pages. Available in PDF, EPUB and Kindle. Book excerpt:

In Vivo Incorporation of Unnatural Amino Acids Into Proteins

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ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (946 download)

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Book Synopsis In Vivo Incorporation of Unnatural Amino Acids Into Proteins by :

Download or read book In Vivo Incorporation of Unnatural Amino Acids Into Proteins written by and published by . This book was released on 2002 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: A method for the site-specific incorporation of unnatural amino acids into proteins in vivo would significantly facilitate studies of the cellular function of proteins, as well as make possible the biosynthesis of unnatural polymers and proteins with novel structures and activities. Our approach consists of the generation of amber suppressor tRNA/aminoacyl-tRNA synthetase pair that are not catalytically competent with all the endogenous Escherichia coli tRNAs an aminoacyl-tRNA synthetases, followed by directed evolution of such orthogonal aminoacyl-tRNA synthetases to alter their amino acid specificities. A new orthogonal suppressor tRNA/aminoacyl-tRNA synthetase pair in E. coli has been derived from the Saccharomyces cerevisiae tRNA (sub Asp) and aspartyl-tRNA syathetase, and the in vitro and in vivo characteristics of this pair were determined. In order to achieve a high specificity for the amino acid, a direct selection for site-specific incorporation of unnatural amino acids into a reporter epitope displayed on the surface of M13 phage has been developed and characterized. Under simulated selection conditions, phage particles displaying aspartate were enriched over 300-fold from a pool of phage displaying asparagine using monoclonal antibodies raised against the aspartate-containing epitope. The direct phage selection offers very high specificity for the amino acid of interest, Which cannot be achieved by conventional methods.

Site-specific Incorporation of Unnatural Amino Acid Into Protein in Vivo

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ISBN 13 :
Total Pages : 506 pages
Book Rating : 4.:/5 (857 download)

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Book Synopsis Site-specific Incorporation of Unnatural Amino Acid Into Protein in Vivo by : Kiket Avi Omer Makoubi

Download or read book Site-specific Incorporation of Unnatural Amino Acid Into Protein in Vivo written by Kiket Avi Omer Makoubi and published by . This book was released on 2012 with total page 506 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Non-Natural Amino Acids

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Publisher : Academic Press
ISBN 13 : 0080921639
Total Pages : 334 pages
Book Rating : 4.0/5 (89 download)

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Book Synopsis Non-Natural Amino Acids by :

Download or read book Non-Natural Amino Acids written by and published by Academic Press. This book was released on 2009-07-24 with total page 334 pages. Available in PDF, EPUB and Kindle. Book excerpt: By combining the tools of organic chemistry with those of physical biochemistry and cell biology, Non-Natural Amino Acids aims to provide fundamental insights into how proteins work within the context of complex biological systems of biomedical interest. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. With more than 400 volumes published, each Methods in Enzymology volume presents material that is relevant in today's labs -- truly an essential publication for researchers in all fields of life sciences. Demonstrates how the tools and principles of chemistry combined with the molecules and processes of living cells can be combined to create molecules with new properties and functions found neither in nature nor in the test tube Presents new insights into the molecular mechanisms of complex biological and chemical systems that can be gained by studying the structure and function of non-natural molecules Provides a "one-stop shop" for tried and tested essential techniques, eliminating the need to wade through untested or unreliable methods

Site Specific Incorporation of Amino Acid Analogues Into Proteins In Vivo

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ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (946 download)

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Book Synopsis Site Specific Incorporation of Amino Acid Analogues Into Proteins In Vivo by :

Download or read book Site Specific Incorporation of Amino Acid Analogues Into Proteins In Vivo written by and published by . This book was released on 2000 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Two critical requirements for developing methods for the site-specific incorporation of amino acid analogues into proteins in vivo are: (1) a suppressor tRNA that is not aminoacylated by any of the endogenous aminoacyl-tRNA synthetases (aaRSs), and (2) an aminoacyl-tRNA synthetase, which aminoacylates the suppressor tRNA but no other tRNA in the cell. Here, we describe two such aaRS/suppressor tRNA pairs, one for use in the yeast S. cerevisiae, and another for use in E. coli. The '21st synthetase/tRNA pairs' include E. coli glutaminyl-tRNA synthetase (GlnRS) along with an amber suppressor derived from human initiator tRNA, for use in yeast, and mutants of the yeast tyrosyl-tRNA synthetase (TyrRS) along with an amber suppressor derived from E. coli initiator tRNA, for use in E. coli. The suppressor tRNAs are aminoacylated in vivo only in the presence of the heterologous aaRSs and the aminoacylated tRNAs function efficiently in suppression of amber codons. Plasmids carrying the E. coli GlnRS gene can be stably maintained in yeast.

Enzyme Engineering

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Publisher : Humana Press
ISBN 13 : 9781493962877
Total Pages : 266 pages
Book Rating : 4.9/5 (628 download)

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Book Synopsis Enzyme Engineering by : James Samuelson

Download or read book Enzyme Engineering written by James Samuelson and published by Humana Press. This book was released on 2016-05-01 with total page 266 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book provides guidance to those wishing to create enzyme variants. It covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes and the engineering of two difference types of rare-cutting endonucleases.

Engineering E. Coli Translational Apparatus

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Publisher :
ISBN 13 :
Total Pages : 416 pages
Book Rating : 4.:/5 (34 download)

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Book Synopsis Engineering E. Coli Translational Apparatus by : Thomas John Magliery

Download or read book Engineering E. Coli Translational Apparatus written by Thomas John Magliery and published by . This book was released on 2001 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Engineering the Genetic Code

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Publisher : John Wiley & Sons
ISBN 13 : 3527607099
Total Pages : 312 pages
Book Rating : 4.5/5 (276 download)

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Book Synopsis Engineering the Genetic Code by : Nediljko Budisa

Download or read book Engineering the Genetic Code written by Nediljko Budisa and published by John Wiley & Sons. This book was released on 2006-05-12 with total page 312 pages. Available in PDF, EPUB and Kindle. Book excerpt: The ability to introduce non-canonical amino acids in vivo has greatly expanded the repertoire of accessible proteins for basic research and biotechnological application. Here, the different methods and strategies to incorporate new or modified amino acids are explained in detail, including a lot of practical advice for first-time users of this powerful technique. Novel applications in protein biochemistry, genomics, biotechnology and biomedicine made possible by the expansion of the genetic code are discussed and numerous examples are given. Essential reading for all molecular life scientists who want to stay ahead in their research.

Exploring and Expanding the Protein Universe with Non-Canonical Amino Acids

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Publisher : Frontiers Media SA
ISBN 13 : 2832538029
Total Pages : 123 pages
Book Rating : 4.8/5 (325 download)

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Book Synopsis Exploring and Expanding the Protein Universe with Non-Canonical Amino Acids by : Gustavo Fuertes

Download or read book Exploring and Expanding the Protein Universe with Non-Canonical Amino Acids written by Gustavo Fuertes and published by Frontiers Media SA. This book was released on 2023-11-01 with total page 123 pages. Available in PDF, EPUB and Kindle. Book excerpt: The site-specific incorporation of unnatural or non-canonical amino acids (ncAAs) into proteins is a universally important tool for systems bioengineering at the interface of chemistry, biology, and biotechnology. The synergistic use of ncAA and related technologies (e.g. Xeno nucleic acids) should enable: i) New opportunities to manipulate, design and elucidate protein structure, dynamics, and function. ii) A deeper understanding of natural and evolved translational systems and their importance for artificial biology. iii) The synthesis of novel biopolymers, creating a solid basis for synthetic cells, which is also an important technology in the production of new classes of medically relevant protein-based scaffolds. Research on reprogrammed protein translation has now reached an experimental and intellectual maturity: more than 200 ncAA (i.e. more than ten times larger variety than standard amino acids) have been introduced into proteins using different routes: genetic code expansion (GCE), selective pressure incorporation (SPI), chemical mutagenesis, protein semi-synthesis, and peptide synthesis.

Site Specific Incorporation of Amino Acid Analogues Into Protiens In Vivo

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ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (946 download)

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Book Synopsis Site Specific Incorporation of Amino Acid Analogues Into Protiens In Vivo by :

Download or read book Site Specific Incorporation of Amino Acid Analogues Into Protiens In Vivo written by and published by . This book was released on 2004 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Our objective is to develop general methods for the site-specific incorporation of amino acid analogues into proteins in bacterial and in eukaryotic cells. The approach consists of the use of an amber suppressor transfer RNA (tRNA) aminoacylated with an amino acid analogue, with the help of a mutant aminoacyl-tRNA synthetase, to insert the amino acid analogue at a specific site in a protein. The site of insertion of the analogue is specified by an appropriately placed amber termination codon within the gene for the protein of interest. This approach has two key requirements: (1) an amber suppressor tRNA, which can not be aminoacylated by any of the endogenous aminoacyltRNA synthetases and (2) an aminoacyi-tRNA synthetase, which aminoacylates the amber suppressor tRNA but no other tRNA in the cell. Therefore, an important first goal is to identify such a 21st aminoacyl-tRNA synthetase tRNA synthetase-amber tRNA pair. This goal has been achieved. Several mutations have been introduced into yeast tyrosyl-tRNA synthetase (TyrRS) for isolating mutants that incorporate iodotyrosine into tRNA instead of tyrosine. Work on an alternative approach applicable to mammalian cells has led to a possible general approach for the introduction of two different amino acid analogues into a protein.

Protein Evolution in the Presence of an Unnatural Amino Acid

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ISBN 13 :
Total Pages : 430 pages
Book Rating : 4.:/5 (871 download)

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Book Synopsis Protein Evolution in the Presence of an Unnatural Amino Acid by : Amrita Singh

Download or read book Protein Evolution in the Presence of an Unnatural Amino Acid written by Amrita Singh and published by . This book was released on 2012 with total page 430 pages. Available in PDF, EPUB and Kindle. Book excerpt: The field of protein engineering has been greatly augmented by the expansion of the genetic code using unnatural amino acids as well as the development of cell-free synthesis systems with high protein yield. Cell-free synthesis systems have improved considerably since they were first described almost 40 years ago. Residue specific incorporation of non-canonical amino acids into proteins is usually performed in vivo using amino acid auxotrophic strains and replacing the natural amino acid with an unnatural amino acid analog. Herein, we present an amino acid depleted cell-free protein synthesis system that can be used to study residue specific replacement of a natural amino acid by an unnatural amino acid analog. This system combines high protein expression yields with a high level of analog substitution in the target protein. To demonstrate the productivity and efficacy of a cell-free synthesis system for residue-specific incorporation of unnatural amino acids in vitro, we use this system to show that 5-fluorotryptophan and 6-fluorotryptophan substituted streptavidin retain the ability to bind biotin despite protein wide replacement of a natural amino acid for the amino acid analog. We envisage this amino acid-depleted cell-free synthesis system being an economical and convenient format for the high-throughput screening of a myriad of amino acid analogs with a variety of protein targets for the study and functional characterization of proteins substituted with unnatural amino acids when compared to the currently employed in vivo format. We use this amino acid depleted cell-free synthesis system for the directed evolution of streptavidin, a protein that finds wide application in molecular biology and biotechnology. We evolve streptavidin using in vitro compartmentalization in emulsions to bind to desthiobiotin and find, at the conclusion of our experiment, that our evolved streptavidin variants are capable of binding to both biotin and desthiobiotin equally well. We also discover a set of mutations for streptavidin that are potentially powerful stabilizing mutations that we believe will be of great use to the greater research community.

A General Method for Site-specific Incorporation of Unnatural Amino Acids Into Proteins

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ISBN 13 :
Total Pages : 7 pages
Book Rating : 4.:/5 (863 download)

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Book Synopsis A General Method for Site-specific Incorporation of Unnatural Amino Acids Into Proteins by : Christopher J. Noren

Download or read book A General Method for Site-specific Incorporation of Unnatural Amino Acids Into Proteins written by Christopher J. Noren and published by . This book was released on 1989 with total page 7 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Protein Engineering

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Publisher : Springer Science & Business Media
ISBN 13 : 354070941X
Total Pages : 351 pages
Book Rating : 4.5/5 (47 download)

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Book Synopsis Protein Engineering by : Caroline Koehrer

Download or read book Protein Engineering written by Caroline Koehrer and published by Springer Science & Business Media. This book was released on 2009-01-07 with total page 351 pages. Available in PDF, EPUB and Kindle. Book excerpt: Site-specific mutagenesis of DNA, developed some thirty years ago, has proven to be one of the most important advances in biology. By allowing the site-specific replacement of any amino acid in a protein with one of the other nineteen amino acids, it ushered in the new era of "Protein Engineering". The field of protein engineering has, however, evolved rapidly since then and the last fifteen years have witnessed remarkable advances through the use of new chemical, biochemical and molecular biological tools towards the synthesis and manipulation of proteins. The chapters included in this book reflect the rapid evolution of protein engineering and its many applications in basic research, biotechnology, material sciences and therapy. This book will provide the reader with an introduction to state-of the-art concepts and methods and will be of use to anyone interested in the study of proteins, in academia as well as in industry.

Protein Engineering

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Publisher : John Wiley & Sons
ISBN 13 : 3527344705
Total Pages : 41 pages
Book Rating : 4.5/5 (273 download)

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Book Synopsis Protein Engineering by : Huimin Zhao

Download or read book Protein Engineering written by Huimin Zhao and published by John Wiley & Sons. This book was released on 2021-08-23 with total page 41 pages. Available in PDF, EPUB and Kindle. Book excerpt: A one-stop reference that reviews protein design strategies to applications in industrial and medical biotechnology Protein Engineering: Tools and Applications is a comprehensive resource that offers a systematic and comprehensive review of the most recent advances in the field, and contains detailed information on the methodologies and strategies behind these approaches. The authors—noted experts on the topic—explore the distinctive advantages and disadvantages of the presented methodologies and strategies in a targeted and focused manner that allows for the adaptation and implementation of the strategies for new applications. The book contains information on the directed evolution, rational design, and semi-rational design of proteins and offers a review of the most recent applications in industrial and medical biotechnology. This important book: Covers technologies and methodologies used in protein engineering Includes the strategies behind the approaches, designed to help with the adaptation and implementation of these strategies for new applications Offers a comprehensive and thorough treatment of protein engineering from primary strategies to applications in industrial and medical biotechnology Presents cutting edge advances in the continuously evolving field of protein engineering Written for students and professionals of bioengineering, biotechnology, biochemistry, Protein Engineering: Tools and Applications offers an essential resource to the design strategies in protein engineering and reviews recent applications.

Noncanonical Amino Acids

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ISBN 13 : 9781493975747
Total Pages : 411 pages
Book Rating : 4.9/5 (757 download)

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Book Synopsis Noncanonical Amino Acids by : Edward A. Lemke

Download or read book Noncanonical Amino Acids written by Edward A. Lemke and published by . This book was released on 2018 with total page 411 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Site-Specific Protein Labeling

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Publisher : Humana Press
ISBN 13 : 9781493922710
Total Pages : 0 pages
Book Rating : 4.9/5 (227 download)

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Book Synopsis Site-Specific Protein Labeling by : Arnaud Gautier

Download or read book Site-Specific Protein Labeling written by Arnaud Gautier and published by Humana Press. This book was released on 2015-01-06 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume provides in-depth protocols for protein labeling techniques and applications, with an additional focus on general background information on the design and generation of the organic molecules used for the labeling step. Chapters provide protocols for labeling techniques and applications, with an additional focus on general background information on the design and generation of the organic molecules used for the labeling step. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Site-Specific Protein Labeling: Methods and Protocols provides a comprehensive overview on the most relevant and established labeling methodologies, and helps researchers to choose the most appropriate labeling method for their biological question.

Chemical and Biological Synthesis

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Publisher : Royal Society of Chemistry
ISBN 13 : 178801507X
Total Pages : 317 pages
Book Rating : 4.7/5 (88 download)

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Book Synopsis Chemical and Biological Synthesis by : Nick J Westwood

Download or read book Chemical and Biological Synthesis written by Nick J Westwood and published by Royal Society of Chemistry. This book was released on 2018-08-16 with total page 317 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synthetic chemistry plays a central role in many areas of chemical biology; utilising recent case studies, the goal of Chemical and Biological Synthesis is to highlight the full impact that the preparation of novel reagents can have in chemical biology. Covering the synthetic approaches that can be applied across the whole field of chemical biology, this book provides synthetic chemists with the broader context to which their work contributes and the biological questions that can be addressed through it. An ideal guide for postgraduate students and researchers in synthetic organic chemistry and chemical biology, Chemical and Biological Synthesis introduces synthetic techniques and methods to those who wish to incorporate synthesis for the first time in their biology-focused research programmes.