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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Murzabaev, Marsel Kojima, Takaaki Mizoguchi, Takuro Kobayashi, Isao DeKosky, Brandon J. Georgiou, George Nakano, Hideo |
| Description | Country affiliation: Japan Author Affiliation: Murzabaev M ( Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.); Kojima T ( Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan. Electronic address: kojimat@nuagr1.agr.nagoya-u.ac.jp.); Mizoguchi T ( Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.); Kobayashi I ( Food Engineering Division, National Food Research Institute, NARO, Tsukuba 305-8642, Japan.); DeKosky BJ ( Department of Chemical Engineering, University of Texas at Austin, Austin, TX 78712, United States.); Georgiou G ( Department of Chemical Engineering, University of Texas at Austin, Austin, TX 78712, United States.); Nakano H ( Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.) |
| Abstract | A simple, inexpensive flow-focusing device has been developed to make uniform droplets for biochemical reactions, such as in vitro transcription and cell-free protein synthesis. The device was fabricated from commercially available components without special equipment. Using the emulsion droplets formed by the device, a class I ligase ribozyme, bcI 23, was successfully synthesized from DNA attached to magnetic microbeads by T7 RNA polymerase. It was also ligated with an RNA substrate on the same microbeads, and detected using flow cytometry with a fluorescent probe. In addition, a single-chain derivative of the lambda Cro protein was expressed using an Escherichia coli cell-free protein synthesis system in emulsion, which was prepared using the flow-focusing device. In both emulsified reactions, usage of the flow-focusing device was able to greatly reduce the coefficient of variation for the amount of RNA or protein displayed on the microbeads, demonstrating the device is advantageous for quantitative analysis in high-throughput screening. |
| File Format | HTM / HTML |
| ISSN | 13891723 |
| Issue Number | 4 |
| Volume Number | 121 |
| e-ISSN | 13474421 |
| Journal | Journal of Bioscience and Bioengineering |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-04-01 |
| Publisher Place | Japan |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Biomedical Engineering Discipline Microbiology Lab-on-a-chip Devices Protein Biosynthesis Transcription, Genetic Cell-free System Dna Genetics Metabolism Dna-directed Rna Polymerases Emulsions Escherichia Coli Flow Cytometry Fluorescence In Vitro Techniques Economics Instrumentation Methods Ligases Analysis Biosynthesis Magnetics Microspheres Rna, Catalytic Repressor Proteins Viral Proteins Viral Regulatory And Accessory Proteins Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Bioengineering Applied Microbiology and Biotechnology Biotechnology |
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