Sunami Takeshi, Kita Hiroshi, Hosoda Kazufumi, Matsuura Tomoaki, Suzuki Hiroaki, Yomo Tetsuya
Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Agency (JST), Chiyoda-ku, Tokyo, Japan.
Methods Enzymol. 2009;464:19-30. doi: 10.1016/S0076-6879(09)64002-7.
In living cells, biochemical reaction systems are enclosed in small lipidic compartments. To experimentally simulate various biochemical reactions occurring in extant cells, giant liposomes are used to reconstruct an artificial model cell. We present methods for conducting a protein synthesis reaction, followed by the reaction catalyzed by the synthesized proteins inside liposomes, and for measurement of the in liposome reaction using a fluorescence-activated cell sorter (FACS). These techniques enable us to perform detailed analysis of the biochemical reactions occurring in the microcompartments, and have the potential to reveal the role of compartmentalization in cellular systems.
在活细胞中，生化反应系统被包裹在小的脂质小室中。为了通过实验模拟现存细胞中发生的各种生化反应，巨型脂质体被用于重建人工模型细胞。我们展示了进行蛋白质合成反应的方法，随后是脂质体内合成蛋白质催化的反应，以及使用荧光激活细胞分选仪（FACS）测量脂质体内反应的方法。这些技术使我们能够对微隔室内发生的生化反应进行详细分析，并有可能揭示细胞系统中分隔化的作用。
