Journal: Nature Communications

Article Title: Self-replication of DNA by its encoded proteins in liposome-based synthetic cells

doi: 10.1038/s41467-018-03926-1

Figure Lengend Snippet: Replication of the Φ29 genome with de novo synthesized proteins. a Reaction pathways for gene expression (IVTT) and DNA amplification. The replication machinery is preferentially directed to the TP-capped Φ29 genome (black arrows). Co-synthesis of the p5 and p6 proteins from their genes is also indicated. b Alkaline agarose gels of the expression-amplification reaction products under various experimental conditions. The p2 and p3 proteins were produced from the oriLR-p2-p3 DNA. The p5 and p6 proteins were expressed from the p5 , p6 , or oriLR-p6-p5 genes. Under these conditions (about equimolar amounts of input Φ29 genome and lower-mass oriLR-p2-p3 DNA), replication is strongly biased toward the natural TP-bound Φ29 genome. The input Φ29 genome can be seen at time zero, while the oriLR-p2-p3 and oriLR-p6-p5 DNAs are visible in some gels (indicated as double and single black asterisks, respectively). The red asterisk indicates the upper band of the Φ29 genome, as also observed with the stock DNA (Supplementary Fig. ) and after amplification by the purified proteins (Supplementary Fig. ). First lane on gels is the DNA ladder. c Quantitative analysis of the experiments shown in b . Values represent the mean and standard deviation (sdv) from three independent experiments. For clarity, only the negative or positive sdv error bars are represented for the full-length product and side products, respectively

Article Snippet: The BenchTop 1-kb DNA Ladder from Promega or the 1-kb Plus DNA ladder from Thermo Fischer Scientific was used to confirm the correct size of the dsDNA templates.

Techniques: Synthesized, Gene Expression, DNA Amplification, Expressing, Amplification, Produced, Purification, Standard Deviation