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We prepared a C-CPE phage library by mutating the functional amino acids.
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The functional consequence of mutating the polyubiquitin binding site of angiocidin was investigated using the Alamar blue viability assay.
We focused on the two regulators, MtrA and MoxR, which were mutated in the functional domain regions in strain E3.
When S188/S190 phosphorylation is prevented by mutating the two residues into alanine, several functional consequences can be observed.
The importance of LBD DBD domain cooperation to assemble the functional complex was demonstrated by mutating the highly conserved Phe375 residue that led to weakened DNA binding.
In the revised manuscript we offer some hints on how the functional consequences from our model might be validated by mutating the acidic residues of Apaf-1.
The functional domains of human GPCRs have over the passed decade been dissected by systematically mutating the peptide sequence [ 8].
That furthered my sense of mutating the genres.
Mutating the 137B site resulted in 16% expression (p<0.0001) which is a little less reduction than was seen when all the potential binding sites were functional.
To further validate that the predicted miR-510 seed sequence within the PRDX1 3'UTR was functional, we mutated the seed sequence of miR-510 in the luciferase reporter construct.
After we had determined the minimum amount of residues required to yield a functional recombinant protein, we mutated the amino acid on the truncation of which the activity was lost, a leucine residue, in a larger construct to investigate whether we simply required a certain length of the protein, or whether specific residues were needed.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com