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We also showed that the distinction between binding sequences and neutrally selected sequences is much stronger in terms of nucleotide content and enrichment.
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To measure the similarity between binding sequences of a pair of factors we assessed the distances between their PSSMs.
We reasoned that if PRDM9 regulates the activity of a significant proportion of hotspots, it should be possible to find a correlation between the predicted binding sequences and recombination rates.
To study the association between the transcription factors and the corresponding binding sequences, and to further confirm the importance of Sp1 and NF-κB in activation of the SPAK gene, we used electrophoretic mobility shift assays (EMSA) to characterize binding of Sp1 and NF-κB to their respective binding sites: Sp1 −496 (Fig. 6A), Sp1 −303 (Fig. 6B), Sp1 −114 (Fig. 6C), and NF-κB −354 (Fig. 6D).
(C ) Relationship between the distance the remodeled nucleosome moved and the separation between the Gal4 binding sequence and the 601NPE.
Instead, the distribution peaked at the midpoint between the Gal4 binding sequence and the 601NPE.
(B ) Distributions of the locations of the nucleosome after ISW1a remodeling in the presence of Gal4DBD on three different templates of increasing separation between the Gal4 binding sequence and the 601NPE.
Thus, the 38 residues between the end of the CALM binding sequence and the start of the transmembrane helix of a VAMP8 molecule must be able to stretch at least this far.
The method, combined with binding sequence and location conservation between human and mouse, identifies with high probability functional binding sites for groups of functionally-related genes.
We used one and two PEG spacers between the binding sequences to fine-tune biological properties of our homodimers.
At the resolution achieved chromosome-wide, we did not find a statistically significant correlation between the predicted binding sequences of the two alleles and recombination rates (Table S5).
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