Figure 4
From: Integrating reductive and synthetic approaches in biology using man-made cell-like compartments
β-Galactoside hydrolysis activity in artificial cell-like compartments.
Multiplex sequencing identified LacZ as a common gene that was included in fluorescent liposomes. To verify this, liposomes that contained the PURE system solution, 100 μM CMFDG, 1 μM transferrin-Alexa Fluor 647 conjugate (volume marker) and 5 nM LacZ were constructed and assayed for their reproducibility of β-galactoside hydrolysis activity. As a negative control, liposomes without LacZ were also constructed. Liposomes were incubated at 37°C and analysed by FACS. At 0 h, CMFDG-derived fluorescence (abscissa) was not detected in either group of liposomes, while after 3 h of incubation, intense fluorescence was detected only in liposomes that contained 5 nM LacZ, which verified the reconstruction of β-galactoside hydrolysis activity. Almost all liposomes had shifted to the right and 8.9% of these liposomes were in the upper right quadrant, which indicated intense fluorescence.
