Tuning relative expression levels of EGFP and mRFP during sequential expression.

<p>Western blot and fluorescence analysis results from monitoring changes in the relative expression levels of the target proteins. NC represents background signal from the reaction that does not contain any template DNA. Magnetic microbead-conjugated plasmids pK7 EGFP and pK7 mRFP were sequentially incubated in the cell-free synthesis reaction mixture using soluble starch as the energy source. With a total incubation time fixed at 180 min, the first template plasmid conjugated on the magnetic microbead was incubated for increasing incubation time periods of 30 min from 30 min to 150 min, followed by incubation of the second plasmid for time period increments of 30 min from 150 min to 30 min. Samples 1 to 5: sequential expression of EGFP and mRFP1, samples 6 to 10: sequential expression of mRFP1 and EGFP.