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Author: John Christopher Anderson
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 546
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Book Description
Several projects focused on expanding the genetic code of Escherichia coli to include amino acids beyond the common twenty are described. These include the identification of extended codons, orthogonal pairs, and the engineering of amino acid metabolic pathways in E. coli . To incorporate multiple unnatural amino acids at distinct sites in a polypeptide, multiple noncoding codons are required. To this end efficiently-suppressed 4- and 5-base codons were identified using a selection strategy. To make use of these extended codons, three orthogonal aminoacyl-tRNA synthetase/suppressor tRNA pairs were engineered from archaebacteria. These systems do not cross-react with their E. coli homologs, but efficiently suppress UAG, AGGA, or UGA codons. Using a combination of saturation mutagenesis and genetic selection, several variants of the Pyrococcus horikoshii lysyl-tRNA synthetase were identified that permit the genetic encoding of homoglutamine in response to the codon AGGA in E. coli . In addition to adding new components to the translational machinery of E. coli , we have also added a biosynthetic pathways for p-aminophenylalanine providing the first example of an autonomous 21-amino acid organism that can biosynthesize an unnatural amino acid and incorporate it into protein with no media supplement. Finally, in an effort to incorporate [alpha]-hydroxy acids genetically into proteins we have identified strains deficient in their biotransformation to [alpha]-amino acids in E. coli . In combination with extended codons and orthogonal pairs, these strains set the stage for the evolution of a system that genetically encodes polyesters. The work described here not only allows us to generate proteins and possibly entire organisms with novel properties not restricted by the common twenty amino acids, it also provides an opportunity to explore basic mechanisms of translation.
Author: John Christopher Anderson
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 546
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Book Description
Several projects focused on expanding the genetic code of Escherichia coli to include amino acids beyond the common twenty are described. These include the identification of extended codons, orthogonal pairs, and the engineering of amino acid metabolic pathways in E. coli . To incorporate multiple unnatural amino acids at distinct sites in a polypeptide, multiple noncoding codons are required. To this end efficiently-suppressed 4- and 5-base codons were identified using a selection strategy. To make use of these extended codons, three orthogonal aminoacyl-tRNA synthetase/suppressor tRNA pairs were engineered from archaebacteria. These systems do not cross-react with their E. coli homologs, but efficiently suppress UAG, AGGA, or UGA codons. Using a combination of saturation mutagenesis and genetic selection, several variants of the Pyrococcus horikoshii lysyl-tRNA synthetase were identified that permit the genetic encoding of homoglutamine in response to the codon AGGA in E. coli . In addition to adding new components to the translational machinery of E. coli , we have also added a biosynthetic pathways for p-aminophenylalanine providing the first example of an autonomous 21-amino acid organism that can biosynthesize an unnatural amino acid and incorporate it into protein with no media supplement. Finally, in an effort to incorporate [alpha]-hydroxy acids genetically into proteins we have identified strains deficient in their biotransformation to [alpha]-amino acids in E. coli . In combination with extended codons and orthogonal pairs, these strains set the stage for the evolution of a system that genetically encodes polyesters. The work described here not only allows us to generate proteins and possibly entire organisms with novel properties not restricted by the common twenty amino acids, it also provides an opportunity to explore basic mechanisms of translation.
Author: Nediljko Budisa
Publisher: John Wiley & Sons
ISBN: 3527607099
Category : Science
Languages : en
Pages : 312
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Book Description
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.
Author:
Publisher: Academic Press
ISBN: 0080921639
Category : Science
Languages : en
Pages : 334
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Book Description
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
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 95
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Book Description
Author:
Publisher:
ISBN: 9780815332183
Category : Cells
Languages : en
Pages : 0
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Book Description
Author: Chang C. Liu
Publisher:
ISBN:
Category : Genetic code
Languages : en
Pages : 438
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Book Description
By expanding the genetic code to allow the cotranslational incorporation of unnatural amino acids, one can site-specifically introduce unique chemical functionality into proteins in vivo. By using such expanded genetic codes in directed evolution, one can impose selection for novel protein function wherein variants containing unnatural amino acids will be selected should the unnatural amino acids confer a functional advantage. I first describe the expansion of E. coli 's genetic code for the site-specific incorporation of the unnatural amino acid sulfotyrosine and the subsequent use of this expanded genetic code for the expression and study of several proteins including sulfo-hirudin and sulfated anti-HIV antibodies. I then describe the development of a phage-based system for evolution with expanded genetic codes and the application of this system for directed evolution, through which proteins that use unnatural amino acids for enhanced function can emerge from selection. This work demonstrates the value of genetically encoding more than the 20 natural amino acids for protein engineering applications and shows that these expanded genetic codes can confer a selective advantage.
Author: Edward A. Lemke
Publisher:
ISBN: 9781493975747
Category : Amino acids
Languages : en
Pages : 411
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Book Description
Author: National Academies of Sciences, Engineering, and Medicine
Publisher: National Academies Press
ISBN: 0309465184
Category : Technology & Engineering
Languages : en
Pages : 189
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Book Description
Scientific advances over the past several decades have accelerated the ability to engineer existing organisms and to potentially create novel ones not found in nature. Synthetic biology, which collectively refers to concepts, approaches, and tools that enable the modification or creation of biological organisms, is being pursued overwhelmingly for beneficial purposes ranging from reducing the burden of disease to improving agricultural yields to remediating pollution. Although the contributions synthetic biology can make in these and other areas hold great promise, it is also possible to imagine malicious uses that could threaten U.S. citizens and military personnel. Making informed decisions about how to address such concerns requires a realistic assessment of the capabilities that could be misused. Biodefense in the Age of Synthetic Biology explores and envisions potential misuses of synthetic biology. This report develops a framework to guide an assessment of the security concerns related to advances in synthetic biology, assesses the levels of concern warranted for such advances, and identifies options that could help mitigate those concerns.
Author: Institute of Medicine
Publisher: National Academies Press
ISBN: 0309219396
Category : Science
Languages : en
Pages : 570
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Book Description
Many potential applications of synthetic and systems biology are relevant to the challenges associated with the detection, surveillance, and responses to emerging and re-emerging infectious diseases. On March 14 and 15, 2011, the Institute of Medicine's (IOM's) Forum on Microbial Threats convened a public workshop in Washington, DC, to explore the current state of the science of synthetic biology, including its dependency on systems biology; discussed the different approaches that scientists are taking to engineer, or reengineer, biological systems; and discussed how the tools and approaches of synthetic and systems biology were being applied to mitigate the risks associated with emerging infectious diseases. The Science and Applications of Synthetic and Systems Biology is organized into sections as a topic-by-topic distillation of the presentations and discussions that took place at the workshop. Its purpose is to present information from relevant experience, to delineate a range of pivotal issues and their respective challenges, and to offer differing perspectives on the topic as discussed and described by the workshop participants. This report also includes a collection of individually authored papers and commentary.
Author: National Academies of Sciences, Engineering, and Medicine
Publisher: National Academies Press
ISBN: 0309437385
Category : Science
Languages : en
Pages : 607
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Book Description
Genetically engineered (GE) crops were first introduced commercially in the 1990s. After two decades of production, some groups and individuals remain critical of the technology based on their concerns about possible adverse effects on human health, the environment, and ethical considerations. At the same time, others are concerned that the technology is not reaching its potential to improve human health and the environment because of stringent regulations and reduced public funding to develop products offering more benefits to society. While the debate about these and other questions related to the genetic engineering techniques of the first 20 years goes on, emerging genetic-engineering technologies are adding new complexities to the conversation. Genetically Engineered Crops builds on previous related Academies reports published between 1987 and 2010 by undertaking a retrospective examination of the purported positive and adverse effects of GE crops and to anticipate what emerging genetic-engineering technologies hold for the future. This report indicates where there are uncertainties about the economic, agronomic, health, safety, or other impacts of GE crops and food, and makes recommendations to fill gaps in safety assessments, increase regulatory clarity, and improve innovations in and access to GE technology.
