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Usually, ChIP is used to test whether specific candidate sequences are bound by a transcription factor, but microarrays are a powerful tool that allows testing large pools of sequences at once.
Using bioinformatics we identified five putative HREs and show that two of these potential regulatory sequences are bound by HIF in vitro and in vivo.
The methods of probe labeling, hybridization, and detection depend on the solid support to which the sequences are bound.
It is therefore possible that some of these sequences are bound by another ETS factor in T cells.
These sequences are bound by proteins of the ATF CREB that play an important role in stress response and that in S. pombe are encoded by the atf1, pcr1, atf21 and atf31 genes [ 46– 46].
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Using the QKI RNA binding consensus sequence, a bioinformatics analysis led to the identification of binding sites in the p27KIP1 (cyclin-dependent kinase inhibitor), MBP and Krox-20 mRNAs and these sequences were bound by the QKI proteins in vitro with high affinity using gel-shift assays [13].
dsDNA sequences were bound to streptavidin-conjugated magnetic beads generating dsDNA probes.
Not all RNA sequences were bound by the RBR to the same extent.
Consensus sequences were bound to the coding sequence using as reference the sequence of Mus musculus retrieved from NCBI GenBank (accession number NM_139301).
The AUC scores are measured by area under ROC curves derived from predictions of gold standard sequences being bound by SRSF1 proteins varying the binding affinity threshold.
Although a consensus HSE (GAAnnTTCnnGAA) has been defined as the ideal high affinity-binding site for HSF1, many variations of this consensus sequence are bound by HSF1 at reduced affinities in vivo.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com