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But it does reproduce itself using the protease and polymerase enzymes.
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Tumors were isolated and sonicated in 0.5% Triton X-100/PBS using the protease inhibitor cocktail (Roche) and analyzed by immunoblotting.
BoLEC and CaLEC were cultured using the protease digestion technique as described elsewhere [ 20].
To this end, we used the protease, reverse transcriptase and integrase domains of each element.
Site mapping was significantly facilitated by using the four proteases and both the phosphoRS score [ 32] and the search engine results, e.g. for four peptides MSPYETHV, MSPYETHVSTDLVGTL, RLMSPYETHVSTDLVGTL and LMSPYETHVSTDLVGTLGYIPPEYGQASVATYK, covering the phosphorylation site Thr (Supplementary Figures S1A S1D respectively).
However, the whole genetic information of A. sojae is insufficient to investigate the functional features important for its industrial use including the protease and amylase activities as well as safety.
dThis value was calculated with biological clones derived from MARLS, using the L (leader) protease-and capsid-coding regions.
The yield and purity of WPs prepared using the two proteases were 16 and 81%% at least, respectively, and the peptides had high quantities (99%% at least) of peptides below 1500 Da with major molecular weight located at 200 1500 Da.
Fluorigenic protease-activity assays were performed using the Enzchek Protease Assay (Invitrogen).
To confirm that the antiserum contains antibodies against the SVSP backbone, the epitope tag was removed from the recombinant protein using AcTEV protease and the resulting protein fragments analysed by immunoblot.
Finally, we cleaved the 6-HIS tag from GFP using TEV protease and tested the proteins for the ability to unfold untagged GFP.
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