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Increase Protein Activity and Yield

ility to synthesize protein with higher biological activity from both prokaryotic and eukaryotic transcripts. Now, more recent data shows ActivePro's ability to synthesize higher yields of multiple proteins in the same reaction. Two viral proteins, protein A and protein B, were synthesized in separate reactions using ActivePro and two other commercial E. coli lysate systems. Figure 1 illustrates the 2-3 fold greater yield of both proteins with ActivePro compared with the competition.

Figure 1. Expression Yields from Two Viral Proteins. DNA templates for protein A and protein B were transcribed/translated in vitro using Ambion's ActivePro Kit (1 hour reaction at 37C) and 2 other commercially available E. coli lysate kits, according to the recommended protocol. Each reaction included 20 Ci 35S-labeled methionine. Yield was assayed by TCA precipitation, and, post-acetone precipitation, 5 l of the reaction was analyzed by SDS-PAGE.


This same translation efficiency also holds true when coexpressing multiple proteins in the same reaction (Figure 2). Cell-free coexpression of two plasmids in the same reaction is useful in interaction studies such as ligand binding experiments, receptor studies, and protein-protein interaction studies. ActivePro has the proven ability not only to coexpress multiple proteins, but to do so with higher yields than the competition.

Figure 2. Cell-free Coexpression of Multiple Proteins. The DNA templates for protein A and protein B (cloned into the included vector) were coexpressed in the same reaction using the ActivePro Kit (1 hour reaction at 37C). The final
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