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It is found that the aptamer I has a higher DNAzyme function (ν = 20.8 µm/min) than that of 1 (ν = 15.3 µm/min).
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The aptamer III is found to exhibit an about two times higher affinity than the aptamer 4. Furthermore, when investigated with the ABTS−H2O2 colorimetry (Figure 6B), the aptamer III also reveals a relatively high DNAzyme function (ν = 9.01 µm/min) as compare with the aptamer 4 (ν = 4.96 µm/min).
The experimental observations show the next generation aptamers all exhibit higher binding affinity and better DNAzyme function than the corresponding original aptamers.
An appropriate base-pairing duplex flanking the G-quadruplex is found to confer higher hemin-binding affinity and more excellent DNAzyme function.
We designed a system in which the DNAzyme function of RNA cleavage was suppressed by the hybridization of an inhibitor strand that possessed disulfide bond with an active DNAzyme.
We achieved a conformational change of oligodeoxynucleotides and the regulation of DNAzyme function by means of a radiolytic strand exchange reaction of disulfide bond.
Also, from the catalytic kinetics of the G-quadruplex-hemin complex over 10 min (Figure 3A), it can be seen that the DNAzyme function of Pu39WT-hemin is almost independent of a K+ or Na+ ion effect.
However, the former exhibits a relatively low DNAzyme function as compared with the latter (Figure 4, curve d).
Interestingly, the aptamer II is also able to bind hemin to form the G-quadruplex-based DNAzyme, and the introduced DNA duplex is found to promote the DNAzyme function of the aptamer II. Figure 4 depicts a comparison between the DNAzyme functions of II and 2 in the ABTS−H2O2 system.
In particular, such G-quadruplex aptamers have another important function, i.e., the peroxidase-like DNAzyme function, which is the emphasis of this study.
That is, the DNAzyme function is improved by about 36% via grafting the DNA duplex onto the G-quadruplex structure of 1.
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