Fig. 2.

Protein synthesis inside giant unilamellar vesicle (GUV) driven by light. Green fluorescent protein (GFP) was synthesized from its messenger RNA (mRNA) (a–e) or DNA (f–h) inside light illuminated GUV (a, c, e–h) or in vitro (b, d). GFP was synthesized inside GUV (a) or in vitro (the PURE system) (b) in which the photosynthesized adenosine triphosphate (ATP) was consumed for the aminoacylation of transfer RNA (tRNA). The insets in a, e and g indicate plot profile of green and red colors on the thin yellow line. c Flow-cytometric analysis of the GUVs of a. The illuminated GUVs are shown as green, whereas the GUVs incubated in the dark are shown as black. The X- and Y-axes represent the fluorescent intensity and the area of forward scattering, respectively. d GFP synthesis coupled with guanosine 5’-triphosphate (GTP) generation. GFP was synthesized in the PURE system with or without nucleoside-diphosphate kinase (NDK), GTP, guanosine diphosphate (GDP) and adenosine 5’-diphosphate (ADP). e The same reactions as in lanes 4 and 6 of d were performed inside GUVs as indicated as NDK+ and NDK−, respectively. f GFP synthesis from its DNA. A gene of whole GFP was introduced in the PURE system with or without bRF_oF₁-PLs, T7 RNA polymerase (T7RNAP), ATP and light. g A small part of GFP (GFP11: 15 amino acids) was synthesized from its encoding DNA inside GUVs containing T7RNAP, another large part of GFP (GFP1-10) purified form E. coli cells, and the PURE system lacking NDK. h The same GUVs of g were analyzed by flow-cytometer as in d. The synthesized GFP in b, d, and f were labeled with [³⁵S]methionine. Scale bar: 10 µm. Source data are provided as a Source Data file