Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes
Directed evolution of proteins is a technique used to modify protein functions through “Darwinian selection.” In vitro compartmentalization (IVC) is an in vitro gene screening system for directed evolution of proteins. IVC establishes the link between genetic information (genotype) and the protein t...
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | Journal of Nucleic Acids |
| Online Access: | http://dx.doi.org/10.1155/2012/923214 |
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| _version_ | 1832545583277015040 |
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| author | Takehiro Nishikawa Takeshi Sunami Tomoaki Matsuura Tetsuya Yomo |
| author_facet | Takehiro Nishikawa Takeshi Sunami Tomoaki Matsuura Tetsuya Yomo |
| author_sort | Takehiro Nishikawa |
| collection | DOAJ |
| description | Directed evolution of proteins is a technique used to modify protein functions through “Darwinian selection.” In vitro compartmentalization (IVC) is an in vitro gene screening system for directed evolution of proteins. IVC establishes the link between genetic information (genotype) and the protein translated from the information (phenotype), which is essential for all directed evolution methods, by encapsulating both in a nonliving microcompartment. Herein, we introduce a new liposome-based IVC system consisting of a liposome, the protein synthesis using recombinant elements (PURE) system and a fluorescence-activated cell sorter (FACS) used as a microcompartment, in vitro protein synthesis system, and high-throughput screen, respectively. Liposome-based IVC is characterized by in vitro protein synthesis from a single copy of a gene in a cell-sized unilamellar liposome and quantitative functional evaluation of the synthesized proteins. Examples of liposome-based IVC for screening proteins such as GFP and β-glucuronidase are described. We discuss the future directions for this method and its applications. |
| format | Article |
| id | doaj-art-7cb832e9474646afa4da12c1d7e58c01 |
| institution | Kabale University |
| issn | 2090-0201 2090-021X |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Nucleic Acids |
| spelling | doaj-art-7cb832e9474646afa4da12c1d7e58c012025-02-03T07:25:21ZengWileyJournal of Nucleic Acids2090-02012090-021X2012-01-01201210.1155/2012/923214923214Directed Evolution of Proteins through In Vitro Protein Synthesis in LiposomesTakehiro Nishikawa0Takeshi Sunami1Tomoaki Matsuura2Tetsuya Yomo3ERATO Japan Science and Technology (JST) and Yomo Dynamical Micro-Scale Reaction Environment Project, Graduate School of Information Sciences and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, JapanERATO Japan Science and Technology (JST) and Yomo Dynamical Micro-Scale Reaction Environment Project, Graduate School of Information Sciences and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, JapanERATO Japan Science and Technology (JST) and Yomo Dynamical Micro-Scale Reaction Environment Project, Graduate School of Information Sciences and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, JapanERATO Japan Science and Technology (JST) and Yomo Dynamical Micro-Scale Reaction Environment Project, Graduate School of Information Sciences and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, JapanDirected evolution of proteins is a technique used to modify protein functions through “Darwinian selection.” In vitro compartmentalization (IVC) is an in vitro gene screening system for directed evolution of proteins. IVC establishes the link between genetic information (genotype) and the protein translated from the information (phenotype), which is essential for all directed evolution methods, by encapsulating both in a nonliving microcompartment. Herein, we introduce a new liposome-based IVC system consisting of a liposome, the protein synthesis using recombinant elements (PURE) system and a fluorescence-activated cell sorter (FACS) used as a microcompartment, in vitro protein synthesis system, and high-throughput screen, respectively. Liposome-based IVC is characterized by in vitro protein synthesis from a single copy of a gene in a cell-sized unilamellar liposome and quantitative functional evaluation of the synthesized proteins. Examples of liposome-based IVC for screening proteins such as GFP and β-glucuronidase are described. We discuss the future directions for this method and its applications.http://dx.doi.org/10.1155/2012/923214 |
| spellingShingle | Takehiro Nishikawa Takeshi Sunami Tomoaki Matsuura Tetsuya Yomo Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes Journal of Nucleic Acids |
| title | Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes |
| title_full | Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes |
| title_fullStr | Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes |
| title_full_unstemmed | Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes |
| title_short | Directed Evolution of Proteins through In Vitro Protein Synthesis in Liposomes |
| title_sort | directed evolution of proteins through in vitro protein synthesis in liposomes |
| url | http://dx.doi.org/10.1155/2012/923214 |
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