The Experts below are selected from a list of 1818489 Experts worldwide ranked by ideXlab platform
Christopher A. Voigt - One of the best experts on this subject based on the ideXlab platform.
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automated design of synthetic ribosome binding sites to control Protein Expression
Nature Biotechnology, 2009Co-Authors: Howard M Salis, Ethan A. Mirsky, Christopher A. VoigtAbstract:Salis et al. design precisely tuned ribosome binding sites that allow rational control over the rate of Protein translation. This technology should facilitate the design of synthetic genetic circuits and metabolic pathways. Microbial engineering often requires fine control over Protein Expression—for example, to connect genetic circuits1,2,3,4,5,6,7 or control flux through a metabolic pathway8,9,10,11,12,13. To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the Protein Expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different Protein Expression levels. We demonstrate the method's utility by rationally optimizing Protein Expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
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Automated design of synthetic ribosome binding sites to control Protein Expression
Nature Biotechnology, 2009Co-Authors: Howard M Salis, Ethan A. Mirsky, Christopher A. VoigtAbstract:Microbial engineering often requires fine control over Protein Expression--for example, to connect genetic circuits or control flux through a metabolic pathway. To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the Protein Expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different Protein Expression levels. We demonstrate the method's utility by rationally optimizing Protein Expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
Howard M Salis - One of the best experts on this subject based on the ideXlab platform.
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automated design of synthetic ribosome binding sites to control Protein Expression
Nature Biotechnology, 2009Co-Authors: Howard M Salis, Ethan A. Mirsky, Christopher A. VoigtAbstract:Salis et al. design precisely tuned ribosome binding sites that allow rational control over the rate of Protein translation. This technology should facilitate the design of synthetic genetic circuits and metabolic pathways. Microbial engineering often requires fine control over Protein Expression—for example, to connect genetic circuits1,2,3,4,5,6,7 or control flux through a metabolic pathway8,9,10,11,12,13. To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the Protein Expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different Protein Expression levels. We demonstrate the method's utility by rationally optimizing Protein Expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
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Automated design of synthetic ribosome binding sites to control Protein Expression
Nature Biotechnology, 2009Co-Authors: Howard M Salis, Ethan A. Mirsky, Christopher A. VoigtAbstract:Microbial engineering often requires fine control over Protein Expression--for example, to connect genetic circuits or control flux through a metabolic pathway. To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the Protein Expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different Protein Expression levels. We demonstrate the method's utility by rationally optimizing Protein Expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
Ethan A. Mirsky - One of the best experts on this subject based on the ideXlab platform.
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automated design of synthetic ribosome binding sites to control Protein Expression
Nature Biotechnology, 2009Co-Authors: Howard M Salis, Ethan A. Mirsky, Christopher A. VoigtAbstract:Salis et al. design precisely tuned ribosome binding sites that allow rational control over the rate of Protein translation. This technology should facilitate the design of synthetic genetic circuits and metabolic pathways. Microbial engineering often requires fine control over Protein Expression—for example, to connect genetic circuits1,2,3,4,5,6,7 or control flux through a metabolic pathway8,9,10,11,12,13. To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the Protein Expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different Protein Expression levels. We demonstrate the method's utility by rationally optimizing Protein Expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
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Automated design of synthetic ribosome binding sites to control Protein Expression
Nature Biotechnology, 2009Co-Authors: Howard M Salis, Ethan A. Mirsky, Christopher A. VoigtAbstract:Microbial engineering often requires fine control over Protein Expression--for example, to connect genetic circuits or control flux through a metabolic pathway. To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the Protein Expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different Protein Expression levels. We demonstrate the method's utility by rationally optimizing Protein Expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
David R. Brown - One of the best experts on this subject based on the ideXlab platform.
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The effects of prion Protein Expression on metal metabolism.
Molecular and cellular neurosciences, 2009Co-Authors: Silvia Kralovicova, Sarah N. Fontaine, Alexandra Alderton, Julia Alderman, K. Vala Ragnarsdottir, Steven J. Collins, David R. BrownAbstract:The prion Protein is a glycoProtein that binds metals such as copper and manganese. When converted to a Proteinase resistant isoform it is associated with prion diseases such as Creutzfeldt-Jakob disease and bovine spongiform encephalopathy. Although, the co-ordination and metal affinity of the prion Protein has been well studied, the association of the Protein with cellular metal metabolism has been less well investigated. We used transgenic manipulation of prion Protein Expression and other recombinant techniques to alter Expression of known copper binding Proteins to investigate the role of the prion Protein in copper metabolism. We found that changing the Expression of the prion Protein alters Proteins associated with copper uptake, storage and export from the cell. In addition, alteration in the Expression of superoxide dismutases increased prion Protein Expression dramatically. Reducing copper in the diet decreased Expression of the prion Protein in the brain while increased dietary manganese dramatically increased the Protein's Expression. Cellular prion infection also increased the Expression of metal transporting Proteins and increased cellular manganese concentrations. Overall our results show a close link between cellular resistance to oxidative stress and also copper metabolism. These findings are in line with previous data suggesting that the prion Protein is an antioxidant and associated with copper uptake into cells. The disturbance to copper metabolism, as a result of altered prion Protein Expression clearly demonstrates the important role of the prion Protein in copper metabolism. The implication is that prion Protein Expression has a homeostatic role in copper metabolism.
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Regulation of prion Protein Expression by noncoding regions of the Prnp gene
Journal of molecular biology, 2007Co-Authors: Cathryn L. Haigh, Josephine A. Wright, David R. BrownAbstract:Expression of the cellular prion Protein is necessary for the transmission and propagation of prion diseases. Increasing the level of prion Protein Expression decreases the incubation period for these diseases. Therefore, understanding the regulation of prion Protein Expression could be critical for treating or preventing these diseases. We investigated the regulation of prion Protein Expression by the promoter and noncoding regions of the bovine and murine Prnp genes. We determined that Expression is modulated by intron 1 and exon 1. In the absence of intron1, exon 1 inhibited activity of the promoter. However, intron 1 demonstrated promoter-like activity and possessed a TATA box. In addition, we identified an alternative transcript present in the brains of cattle and mice that lacks exon 1. Taken together, these results show that intron 1 and exon 1 play a critical role in the regulation of prion Protein Expression. Because switching off prion Protein Expression has been shown to arrest prion disease, these regions present novel targets for intervention in the disease process.
Kenji Miura - One of the best experts on this subject based on the ideXlab platform.
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Agroinfiltration-based efficient transient Protein Expression in leguminous plants
Plant Biotechnology, 2019Co-Authors: Takuya Suzaki, Mai Tsuda, Hiroshi Ezura, Kenji MiuraAbstract:Transient Protein Expression is an effective tool to rapidly unravel novel gene functions, such as transcriptional activity of promoters and sub-cellular localization of Proteins. However, transient Expression is not applicable to some species and varieties because of insufficient Expression levels. We recently developed one of the strongest agroinfiltration-based transient Protein Expression systems for plant cells, termed 'Tsukuba system.' About 4 mg/g fresh weight of Protein Expression in Nicotiana benthamiana was obtained using this system. The vector pBYR2HS, which contains a geminiviral replication system and a double terminator, can be used in various plant species and varieties, including lettuces, eggplants, tomatoes, hot peppers, and orchids. In this study, we assessed the applicability of the Tsukuba system to several species of legumes, including Lotus japonicus, soybean Glycine max, and common bean Phaseolus vulgaris. The GFP Protein was transiently expressed in the seedpods of all examined legume species, however, Protein Expression in leaves was observed only in P. vulgaris. Taken together, our system is an effective tool to examine gene function rapidly in several legume species based on transient Protein Expression.
