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In contrast, the sequences of the EPIYA-C type differ from the SHP-2 binding sequence by a single amino acid located in the pY + 5th position [ 51, 52].
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While it is easy to create a single binding sequence by point mutation, the appearance of a combination of several binding motifs in an extended region of DNA is definitely more unlikely in this way.
The rationale behind this study is that while it is easy to create a single binding sequence by point mutation it is difficult to understand how a local evolutionary process could create an extended (non-local) combination of binding sequences.
For this experiment, we designed a long template (∼1200 bp) with a 601NPE separated from a Gal4 binding sequence by 11 bp.
Two reported TGFβ1 binding peptides were synthesized with a terminal cysteine separated from the binding sequence by two glycine spacers (CGG WSHW(29) and CGG KRIWFIRPSSWY 30)) and then covalently linked to a dextran-functionalized SPR flowcell surface using standard ligand-thiol coupling chemistry.
Further, the selected antigenic sequences (having B-cell epitopes) were analyzed for T-cell epitopes (MHC I and MHC II alleles binding sequence) by using ProPred 1 and ProPred respectively.
For templates used in (D )–(G ), the 601NPE was separated from the Gal4 binding sequence by 10 bp.
By having all required data at hand we can re-adjust inaccurately determined TF binding sequences by shifting some motifs by some positions and by assigning a strand annotation, if necessary.
To validate these predicted genes, we searched the GCN4 motif for known OsSMF1 binding sequences by combined microarray analysis.
The present study focused only on one level of TF-DNA interaction, the recognition mechanisms of DNA binding sequences by transcription factors.
Next, we calculated the 'match score' of these 115 AdpA-binding sequences to the consensus AdpA-binding sequence by Mast and examined a relationship between the match score (which represented the degree of similarity to the AdpA-binding consensus sequence) and AdpA-binding strength.
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