用于酶促反应的微生化反应器,包括无细胞mRNA翻译

T. Fujii, S. Shoji, T. Nojima, A. Yotsumoto, K. Hosokawa, I. Endo
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引用次数: 5

摘要

作为迈向“芯片上的实验室”或“芯片上的细胞”概念的第一步,使用传统的MEMS(微机电系统)技术制造了几个微生化反应器:硅各向异性蚀刻和玻璃硅阳极键合。微反应器主反应通道长14 mm,宽200 ~ 800 μm,深20 μm。在这些通道中,酶促反应-荧光素-荧光素酶反应和无细胞mRNA翻译-被证实。荧光素-荧光素酶反应诱导的发光以两种方式观察,一种是高速胶片照相,另一种是灵敏视频成像。在这两种情况下,微反应器中基于扩散的混合都清晰可见。在无细胞蛋白合成实验中,采用放射性同位素法测定多尿苷酸模板在微反应器中翻译合成的聚苯丙氨酸的量。
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Microbiochemical reactors for enzymatic reactions including cell-free mRNA translation
As the first step toward the 'lab on a chip' or 'cell on a chip' concept, several microbiochemical reactors were fabricated using conventional MEMS (microelectromechanical systems) techniques: silicon anisotropic etching and glass-silicon anodic bonding. The microreactors have main reaction channels which are 14 mm long, 200-800 μm wide, and 20 μm deep. In these channels, enzymatic reactions - luciferin-luciferase reaction and cell-free mRNA translation - were demonstrated. Luminescence induced by the luciferin-luciferase reaction was observed in two ways, by photograph on a high-speed film and sensitive video imaging. In both cases, diffusion-based mixing in the microreactors was clearly visualized. In the cell-free protein synthesis experiment, the amount of polyphenylalanine, which was synthesized by translating polyuridylic acid template in the microreactors, was determined using radioisotope assay.
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