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Translate More Protein with Higher Biological Activity

ein per reaction. Rhodanese (eukaryotic) or -galactosidase (prokaryotic) proteins were synthesized using ActivePro and two other commercial E.coli lysate systems. The reactions were then used in a functional assay specific for each protein to measure total protein activity (units of activity/ml reaction). Figure 3 demonstrates that ActivePro generates proteins with higher activity than other kits.

Figure 3. Biological Activity of Proteins Produced Using the ActivePro Kit vs. Other Commercial Kits. DNA template (1 g) for -galactosidase (-gal) (A) or rhodanese (B) was used in either an ActivePro Kit reaction or other commercially available E.coli lysate kits according to protocol. The reaction (1 l) was assayed for -gal by colorimetric ONGP hydrolysis assay and rhodanese by the colorimetric assay as described by Sorbo1.


Proteins synthesized using ActivePro also show higher specific activity (expressed as activity units/mg protein) when compared to proteins translated in other systems. Figure 4 shows that the specific activity of proteins made with ActivePro was much higher than Kit 1 (a commercially available kit) and may even approach or equal the activity of the native protein (Figure 3, Panel A). Figure 5 demonstrates the sharp bands produced by ActivePro translation, suggesting that full-length translation products are produced. An increase in specific activity indicates an increase in full length active protein. Proteins with high specific activity increase the availability of functional protein for downstream assays. Even though other kits produced high protein yields, these proteins did not show high specific acti
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