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In this paper, we present an idea to inhibit the evolving receptors by using an ensemble of all possible single- or double-point mutant sequences of the ligand peptide.
We compared λmaxof 12 groups (Fig. 3), statistically significant differences were observed between wildtype and mutant sequences of all groups (P < 0.05, Student'st- test for paired data).
The two mutant sequences of miR-124 were designed by switching bases at positions 5 and 6 (124mut5-6) and at positions 9 and 10 (124mut9-10), respectively.
In their experimental work, Moldoveanu et al. were able to induce proteolysis in several mutant sequences of calpastatin by deleting one or two specific residues (Lys176, Glu177, or both) from a sequence of wild type rat calpastatin (gi 13540322) [49].
In contrast, in cells transfected with plasmids containing mutant sequences of the PDCD4 3′-UTR, luciferactivityvity was minimally decreased in TE13 cells and the reporter activity in Eca109 cells was even increased.
Then, we used a surface plasmon resonance (SPR) biosensor to test 3 mutant sequences of high ZRANK scores along with a high and a low affinity binding sequence as reported in the literature.
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The mutant sequence of cloned PCR products was confirmed by DNA sequencing.
The proposed biosensor can distinguish between the normal sequence and the mutant sequence of ApoB-100 gene, promising a possibility to apply the QD-based biosensor for clinical investigations.
The surviving yeast consisted of RTSH yeast (the mutant sequence of CBM: N18R, S23T, S26S, and T27H) and wild-type yeast (CBM was the original) in a ratio of 1 46.
Based on our findings, we prepared the reporter gene assay with wild or mutant sequence of the hypothetical binding site of TNRC6B and miR-18b.
This investigation presents a new in vitro SNP discrimination mechanism, demonstrated by discriminating between the wild-type and mutant sequence of the mitochondrial A8344G MERRF mutation.
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