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The location of the obtained motifs in the tested gene set promoters was visualized by the Motif Align and Search Tool (MAST) within the MEME software suite [48].
Veerla et al. recently developed SMART software for identifying co-occurring TFBSs in gene set promoters [ 7].
For this reason, we re-cloned the nine MA data set promoters using the ATG translation start site of each gene; three putative BD PROs that displayed bidirectional activity and five that showed promoter activity in one direction were recovered (MA-n without a red box in Figure 3A).
We also re-cloned the 18 RS data set promoters using the ATG start site of each gene and recovered 11 putative BD PROs that functioned as BD PROs and 7 that showed promoter activity in one direction (RS-n without a red box in Figure 3B).
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Using this method to edit the GAL1 and GAL80 promoter sequences, we found that the relative positioning of promoter elements was critically important for setting promoter activity levels in single cells.
Moreover, the number of c-Myc binding sites identified in the promoter set including promoters of induced genes was compared to the number of c-Myc binding sites identified in the control promoter set.
The number of transcription factor binding sites identified in the analyzed promoter set was compared to the number of transcription factor binding sites identified in a control promoter set [promoters of 100 selected genes which were not regulated at all in all four different groups].
As co-MYC regions showed a strong tendency to be located close to annotated TSSs we termed this enhancer negative training set promoter-like (PrL, see Methods).
We produced the "universe" set of promoters for a given epigenetic mark by forming, for each epigenetic mark, the union of positive promoters across BTSC types.
The latter sets deviate from a Poisson distribution (12 species hsp70: chi-square, p<0.0001, Hsp70 in D. melanogaster natural populations: p<0.0001; Set I promoters in D. melanogaster natural populations, p<0.0001), whereas those inserting into the genesets for the 12 species do not (Promoter set I: p = 0.743, Promoter set II: p = 0.279, Promoter set III: p = 0.272).
This can happen, for example, when the small input set of promoters all happen to be AT rich.
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