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We then count the matches between this set of footprints and the binding regions from a MacIsaac gold standard.
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Removal of the Rbf1 binding region from an InR reporter inhibited this repression by Rbf1 (Y. Wei, unpublished data).
Structural modelling of AbrB bound to the target sequence indicated that the AbrB tetramer would interact with ∼20 bp sequences, whereas in vitro footprinting studies detected a wider range of binding regions from 25 to 80 bp, suggesting that a higher order structure of the AbrB tetramer may be involved in DNA binding at some sites on the chromosome.
We performed the same experiment on 11 SUZ12 binding regions from ChIP-seq data and similarly found a strong correlation (r2 = 0.792, Figure S1).
Previous structural modelling of AbrB bound to the target DNA sequence indicated that the AbrB tetramer would interact with ∼20 bp sequences, whereas in vitro footprinting studies detected a wider range of binding regions from 25to8080 bp.
In this study, we propose a computational approach to refine the detection of protein binding regions from pyro-sequencing-derived RNA fragments.
Based on a traditional EMSA and selection of DNA fragments containing protein binding regions from the population of restriction fragments (similar to the SELEX methodology [ 13]), we have developed a strategy for the detection of protein binding regions in long (<10 kb) fragments of genomic DNA (Fig. 1).
Additionally, different membrane binding regions from membrane fusion proteins are necessary to complete the fusion process [34].
To verify this hypothesis, we randomly picked 28 Nanog ChIP-seq detected binding regions from [35] and repeated the ChIP experiments in E14 mouse ES cells.
PSSMs were used to scan 13720 described CTCF binding regions from −500 bp to +500 bp relative to individual peak centers using the Patser tool of the RSAT suite [22].
This feature allows us to refine the detection of protein binding regions from pyro-sequencing-derived RNA fragments.
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