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For molecular functions, binding activity was the most represented GO term, followed by catalytic activity.
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The smallest format found to keep full binding activity is the so-called single-chain fragment (scFv), in which the two variable regions, VH and VL, are artificially linked by a flexible polypeptide [12] [15].
Catalytic activity and binding activity were the primary molecular functions associated with BDTPs.
The three most highly enriched GO terms for Cellular Component were membrane, intracellular and cytoplasm, while transferase activity, hydrolase activity and nucleotide binding activity were the most enriched in the Molecular Function category.
Among three GO categories, cell and binding activity were the most abundant classes in cellular component and molecular function, respectively, which was consistent with the report from Xia et al. [ 24].
A second requirement for repeats 9 and 10 to fully express their fibronectin-binding activity was the non-binding repeat 11.
Regarding MF GO categories (Additional file 2: Figure S2A and S3A), the catalytic and binding activities were the most represented subcategories for both up- and downregulated GO terms.
The RNA binding activity was assayed using the similar method but the labeled RNA oligonucleotide with the sequence 13 mer 5′-AUAGCCUCAACC-G-3′ used for making the RNA substrate was used for the assay.
The same binding activity was detected for the region in front of the glcD gene, whereas the complex with the predicted binding site in front of the mraZ gene was much weaker.
PAMPs binding assay was performed by the method of PAMPs microarrays, and the binding activity was calculated by the signal value of fluorescence.
To verify that the binding activity was due to the presence of Pax-5 we performed a super-shift experiment using a Pax-5 specific antibody, a control antibody or no antibody in the binding reaction.
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