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Mutations designed in the C-terminal flanking region of the PDZ domain resulted in a significant decrease in binding affinity for E6 peptides.
The two mutations designed in S9 included either a half-deletion of the first flexible loop of VP6 (nt 115 nt 291) (d1), or a whole deletion (nt 115 nt 405) (d2) (Fig. 3A).
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Using an in vivo minigenome replication system, we engineered mutations designed to disrupt potential base pairing in the viral RNA termini.
Two steps were followed to study the activation mechanism of CheY upon phosphorylation: first, we independently substituted the three aspartic acid residues in the active site with alanine; second, several mutations were designed in helix α4, both to increase its level of stability and to improve its packing against the protein core.
A first group of mutations were designed in order to disrupt putative interactions between the agonist and the binding pocket (Figure S3).
The K2A mutation was designed in light of the strong tendency of the N-terminus region to interact with sequestered PIP2 lipids, as observed in the MD simulations of STX in PM membranes.
A series of mutation assays were designed in order to confirm these proposed contact regions and identify the specific interaction sites.
The patented bird carries a genetic mutation designed to suppress the "white gene" in baby turkeys and then allow it to emerge in adult birds.
As always, the specific sexual reproduction and mutation rules should be designed in a manner which ensures that their result encodes a valid solution to the problem at hand.
Therefore, the double mutations T166A/E539K and K312E/E539K were designed in the hope of obtaining mutants with higher catalytic efficiency and better thermostability.
Combining one of these cavity mutations with the best variant designed in our previous study led to a sevenfold increase in affinity compared to the wild-type receptor.
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