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Among the mutants tested were a type III secretion mutant (pscD), mutants in three different type III virulence effectors (ExoT, ExoU, and ExoY), an exoTUY triple mutant, a mutant in a secreted exotoxin (toxA), and two mutants in a gene involved in the production of toxic hydrogen cyanide (hcnC).
Mutants in three such genes (bus-10, srf-2, and srf-5 ) were used initially.
To further illustrate the power of DASH to resolve closely migrating oligosaccharides, we characterised further mutants in three closely related proteins in the DUF579 family.
At least for us, it is impossible to evaluate such a large number of mutants in three different assay systems (in vitro phosphorylation, oocyte experiments, and mutant complementation).
Using identical alleles, we compared multiple SUN domain mutants in three commonly used S. cerevisiae strain backgrounds: W303, BY (a S288c derivative), and SK-1.
Hence, YEN1/GEN1 mutants in three different organisms lack the phenotypes predicted for the major HJ resolvase acting in HR based on the classic model for HJ formation as a major recombination intermediate.
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Insertion mutants in two genes with yet unknown function, however, were found to attain significantly improved riboflavin titers and yields.
These are used to demonstrate that an active variant can be selected from a background of 10,000 inactive mutants in four rounds of selection and amplification.
First, we considered a training database of 339 mutants in nine different proteins and optimised the set of parameters and weighting factors that best accounted for the changes in stability of the mutants.
Here, we systematically investigate the behavior of these disease mutants in four different cell types: three not containing desmin or the related IF protein vimentin and the standard fibroblast line 3T3, which has an extensive vimentin system.
Complementation failed to restore the S phenotype in mutants in four of these ORFs.
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CEO of Professional Science Editing for Scientists @ prosciediting.com