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Structural information and an understanding of how proteins respond to mutation and recombination are being used to develop improved directed evolution strategies by increasing the probability that mutant sequences have the desired properties.
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The mutant cDNA sequences have been digested with EcoRI and NotI restriction enzymes, to be cloned into a previously digested pCDNA5/FRT.
Nevertheless, not all mutant sequences could have a high PSA-binding ability.
The Tos17 mutant collection consisting of approximately 50,000 mutant lines characterized into flanking sequences have been used in full characterization of many genes characterized in Japan and nearly 50 Tos17 mutant lines have been successfully used in tagging specific genes (See reference list at https://tos.nias.affrc.go.jp/doc/references.html).html
It should be noticed that the three mutant sequences all had smaller dissociation rate constants.
In the case of N-terminal mutants of A-RAF, BamHI and HindIII recognition sequences have been attached to the sequence of forward and reverse primers respectively.
Notably, a mutant CXCL12 sequence has been designed that has minimal affinity for HS (18).
The correlation is not strictly linear since, for example, the mutant sequence 8mUB1 has almost the same base-pair probability value as 3mUB1 and 5mUB1, although it is more efficiently spliced than these two.
Mutants to be sequenced have been selected on the basis of their fabI promoter sequence in order to analyse one strain for each one of the previously identified mutations.
Indeed, assessment of the transactivation capacity of p53 mutant proteins on different p53 target sequences has shown great variability of activities between mutant proteins and target sequences.
Sequences having a mixture of wild-type and mutant residues at single positions were considered to have the mutant(s) at that position.
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