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Fig. 3 | BMC Biology

Fig. 3

From: Sequence-based prediction of permissive stretches for internal protein tagging and knockdown

Fig. 3

Cleavability of internally TEV-tagged protein variants. Cleavability of TEV-tagged Adk-6xHis variants was examined by incubating crude lysates derived from strains expressing the indicated Adk variant from a low copy vector under control of the natural Adk promoter in the presence or absence of TEV protease. Samples were separated by SDS PAGE and blotted. Cleavage products were detected with a 6xHis-tag-specific antibody. a AdkD76.1, b AdkA99, c AdkA186. The predicted secondary structure context of each residue is indicated in red. The loop length for variants AdkD76 and A99 was determined based on secondary structure prediction. d Cleavage of flanked TEV-tag variants inserted after residue D76. Purified AdkD76-Strep variants with differing TEV-tag flanking region lengths were incubated in the presence or absence of TEV protease and cleavage products were detected with a Strep-tag-specific antibody

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