Journal: Nature Communications

Article Title: Investigation of artificial cells containing the Par system for bacterial plasmid segregation and inheritance mimicry

doi: 10.1038/s41467-024-49412-9

Figure Lengend Snippet: a Giant unilamellar vesicles (GUV) containing the ParMRC system and PURE system. b parC -eGFP DNA segregation by the polymerization of ParM triggered by adenosine triphosphate (ATP) influx upon laser irradiation. c A deformed GUV under hypertonic conditions. d Two daughter GUVs containing parC -eGFP DNA and the PURE system upon laser irradiation at the center region of the deformed GUV (purple area in Fig. 1c). e Enhanced green fluorescent protein (eGFP) was expressed inside two daughter GUVs through translating the eGFP gene using a PURE (protein synthesis using recombinant elements) system at 37 °C. The PURE system contains ribosomes, amino acids, nucleoside triphosphates (NTPs), transfer ribonucleic acid (tRNAs), enzyme substrates, RNA polymerase, translation factors, and other necessary components.

Article Snippet: A Bradford protein assay kit, SDS‒PAGE gel configuration kit, 4×SDS‒PAGE sample loading buffer, protease inhibitor cocktail for purification of His-tagged proteins, green fluorescent DNA marker dye, streptavidin magnetic beads, Tris-borate-EDTA buffer (TBE) and Tween-20 were purchased from Beyotime (China).

Techniques: Irradiation, Recombinant

Journal: Nature Communications

Article Title: Investigation of artificial cells containing the Par system for bacterial plasmid segregation and inheritance mimicry

doi: 10.1038/s41467-024-49412-9

Figure Lengend Snippet: a Schematic illustration of two parC -eGFP DNAs being pushed through ParM polymerization in a giant unilamellar vesicles (GUV) and subsequent GUV division under laser irradiation. b Confocal microscopy images of GUV division ( b1 – b3 ) and enhanced green fluorescent protein (eGFP) expression at 37 °C ( b4 – b7 ). b1 – b3 indicate GUV deformation, filament splitting, and division into two daughter cells, respectively. The white arrows in b1 and b2 indicate parC -eGFP DNA. The ParM filament was split by laser irradiation (561 nm, 0.7 mW, 5 s). The scale bars are 10 μm. n = 3 independent replicates. c Schematic illustration of eGFP expression in two daughter cells. The corresponding fluorescence intensity (FI) in daughter GUV 1 ( d ) and daughter GUV 2 ( e ) as a function of time. The normalized fluorescence intensity of eGFP was calculated from three independent samples. The data were presented as the mean values ± SDs; n = 3 independent replicates. Source data are provided as a Source Data file.

Article Snippet: A Bradford protein assay kit, SDS‒PAGE gel configuration kit, 4×SDS‒PAGE sample loading buffer, protease inhibitor cocktail for purification of His-tagged proteins, green fluorescent DNA marker dye, streptavidin magnetic beads, Tris-borate-EDTA buffer (TBE) and Tween-20 were purchased from Beyotime (China).

Techniques: Irradiation, Confocal Microscopy, Expressing, Fluorescence