Multiphase water-in-oil emulsion droplets for cell-free transcription-translation.
Multiphase water-in-oil emulsion droplets for cell-free transcription-translation.
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DOI：
10.1021/la404146g
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发表时间：
2014-05-27
期刊：
影响因子：
--
通讯作者：
Mansy SS
中科院分区：
文献类型：
--
作者：
Torre P;Keating CD;Mansy SS
在含有有组织的合成胞质溶胶的隔室中构建遗传编码的细胞模拟物对于人工细胞的开发是期望的。以与转录和翻译机制相容的方式将相分离的水性结构域置于油包水乳液液滴内。水两相和三相系统（ATPS和A3PS）与葡聚糖，聚（乙二醇），和Ficoll组装。含水两相系统能够支持水滴内的蛋白质的无细胞表达，而基于含水三相的系统没有引起可检测的蛋白质合成。表达的蛋白优先分配到葡聚糖富集相。该系统可以作为构建具有液体细胞器的细胞模拟物的基础。
The construction of genetically encoded cellular mimics in compartments containing organized synthetic cytosols is desirable for the development of artificial cells. Phase separated aqueous domains were placed within water-in-oil emulsion droplets in a manner compatible with transcription and translation machinery. Aqueous two-phase and three-phase systems (ATPS and A3PS) were assembled with dextran, poly(ethylene glycol), and Ficoll. Aqueous two-phase systems were capable of supporting the cell-free expression of protein within water droplets, whereas the aqueous three-phase-based system did not give rise to detectable protein synthesis. The expressed protein preferentially partitioned to the dextran-enriched phase. The system could serve as a foundation for building cellular mimics with liquid organelles.
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