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Synthesis of milligram quantities of proteins using a reconstituted in vitro protein synthesis system.
Kazuta, Yasuaki; Matsuura, Tomoaki; Ichihashi, Norikazu; Yomo, Tetsuya.
Afiliación
  • Kazuta Y; Japan Science and Technology (JST), ERATO, Dynamical Microscale Reaction Environment Project, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan.
  • Matsuura T; Japan Science and Technology (JST), ERATO, Dynamical Microscale Reaction Environment Project, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan; Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
  • Ichihashi N; Japan Science and Technology (JST), ERATO, Dynamical Microscale Reaction Environment Project, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan.
  • Yomo T; Japan Science and Technology (JST), ERATO, Dynamical Microscale Reaction Environment Project, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan; Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka 565-0871, Japan; Graduate School of Frontier Biosciences, Os
J Biosci Bioeng ; 118(5): 554-7, 2014 Nov.
Article en En | MEDLINE | ID: mdl-24880499
In this study, the amount of protein synthesized using an in vitro protein synthesis system composed of only highly purified components (the PURE system) was optimized. By varying the concentrations of each system component, we determined the component concentrations that result in the synthesis of 0.38 mg/mL green fluorescent protein (GFP) in batch mode and 3.8 mg/mL GFP in dialysis mode. In dialysis mode, protein concentrations of 4.3 and 4.4 mg/mL were synthesized for dihydrofolate reductase and ß-galactosidase, respectively. Using the optimized system, the synthesized protein represented 30% (w/w) of the total protein, which is comparable to the level of overexpressed protein in Escherichia coli cells. This optimized reconstituted in vitro protein synthesis system may potentially be useful for various applications, including in vitro directed evolution of proteins, artificial cell assembly, and protein structural studies.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Biosíntesis de Proteínas / Técnicas In Vitro Idioma: En Revista: J Biosci Bioeng Asunto de la revista: ENGENHARIA BIOMEDICA / MICROBIOLOGIA Año: 2014 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Japón
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Biosíntesis de Proteínas / Técnicas In Vitro Idioma: En Revista: J Biosci Bioeng Asunto de la revista: ENGENHARIA BIOMEDICA / MICROBIOLOGIA Año: 2014 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Japón