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Since the small GTPases Rac1 and Cdc42 share 66.8% of DNA sequence identity, we controlled the specificity of the designed shRNA towards Cdc42 by comparing its action on the expression level of both Cdc42 and Rac1.
We controlled the specificity of the assay: i) by using a siRNA knocking down another eukaryotic protein, histone H1, which has been associated with many effects of nucleolin [14], [15]; and ii) by monitoring entry of either another intracellular pathogen with a cytosolic niche, Listeria monocytogenes, or of fluorescent beads, with the same average 0.5 µm diameter as F. tularensis.
The –e 100 parameter specified the expectation value cutoff and controlled the specificity.
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To control the specificity of the HLA-A2/IDO5 tetramer we stained an IDO5-specific T-cell clone.
However, the mechanism controlling the specificity of the SMYD2 ERα interaction and the structural basis of SMYD2 substrate binding diversity are unknown.
Protein engineering is becoming increasingly important for pharmaceutical applications where controlling the specificity and affinity of engineered proteins is required to create targeted protein therapeutics.
A key component of the enzyme cascade that results in attachment of the multi-ubiquitin chain to the target or labile protein is the ubiquitin ligase that controls the specificity of the ubiquitination reaction.
There are two main families of modern tools for genome editing depending on the molecule that controls the specificity of the system and drives the editor machinery to its place of action.
(A) Negative control (NC) omitting the primary sGC antibody to control the specificity of the immunostaining with Alexa 488.
To further control the specificity of induced phenotypes in Eph4ΔC expressing cells, two other mutant forms of Eph, Eph4TM and Eph3ΔC, were used.
The DNA binding domain of c-Myb is highly conserved amongst vertebrates and even insects and plants [34], [35], and is presumed to control the specificity of c-Myb by recognizing and forming stable complexes at the appropriate promoters.
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