We sought to prepare millimeter-sized supergiant unilamellar vesicles (SGUVs) by spontaneous emulsion transfer for efficient, eukaryotic cell-free translation in the interior. Although the conventional protocols require that a considerably high concentration of sucrose be encapsulated into the SGUVs for their efficient formation, such high amounts of sucrose severely inhibited cell-free translation based on wheat germ extract (WGE). We thus optimized the preparation conditions to permit SGUV formation at a much lower concentration of sucrose that has almost no effect on WGE translation. Under the optimized conditions, we successfully prepared WGE translation system-encapsulating SGUVs that allow for protein synthesis with a high efficiency comparable to that outside a liposome. The optimization also resulted in a high rate of successful SGUV formation (>90%) and a decent stability of the formed SGUVs (>60 min). These SGUVs are expected to serve as research tools in cell-free synthetic biology and as foundations for artificial cell-based biosensors.

中文翻译：

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毫米大小的超大型脂质体的制备，该脂质体可通过自发的乳液转移在内部进行高效，真核的无细胞翻译。

我们试图通过自发乳剂转移制备毫米大小的超大型单层囊泡（SGUV），以在内部进行高效，真核的无细胞翻译。尽管常规方案要求将相当高浓度的蔗糖封装到SGUV中以有效形成，但是基于小麦胚芽提取物（WGE），如此大量的蔗糖严重抑制了无细胞翻译。因此，我们优化了制备条件，以允许以低得多的蔗糖浓度形成SGUV，而这几乎对WGE翻译没有影响。在最优化的条件下，我们成功地制备了WGE翻译系统，其包封了SGUV，可实现与脂质体外部相当的高效蛋白质合成。优化还导致成功完成SGUV的比率很高（> 90％），并且所形成的SGUV的稳定性也很好（> 60分钟）。这些SGUV有望作为无细胞合成生物学的研究工具，并为基于人工细胞的生物传感器奠定基础。