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Surprisingly the resulting strain grew anaerobically on xylose in synthetic media with a μmax as high as 0.09 h−1 without any non-defined mutagenesis or selection.
All lesions were introduced by transduction, site-directed mutagenesis, or selection of spontaneous mutations.
The localized high mutation frequency correlated with restoration of DMS4 function implies an efficient mechanism for targeted mutagenesis or selection of more fit revertant cells in the shoot apical meristem, thereby rapidly restoring a wild-type phenotype that is transmitted to future generations.
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We discuss this unexpectedly high mutation frequency in the context of targeted mutagenesis of the dms4 gene or selection of more fit revertant cells in the shoot apical meristem.
These hypothetical mechanisms can also be applied to our results by positing targeted mutagenesis of the dms4-1 allele or selection of revertant cells containing a dms4-1 suppressor mutation in the SAM.
For the site-directed or site-saturation mutagenesis, the selection of the amino acid positions or the substituted residues was a pivotal step, which could be predicted based on in silico design or computer-aided design (Yin et al. 2015).
Comparable cefotaxime resistant variants were selected relatively quickly (only two rounds of mutagenesis and selection) and with no silent or other superfluous mutations, an approach that was accomplished through what we have termed a "minimal mutational pathway".
Since the viral genome can be carried as an episome in bacterial cells, prokaryotic genetic techniques can be employed for manipulation of the viral genome without the need for typical selective mechanisms required for mutagenesis and selection in eukaryotic cells.
An industrially important cellulolytic strain of T. reesei, Rut-C30, was selected for improved cellulase production following multiple rounds of mutagenesis and selection.
Consecutive mutagenesis and selection using display systems that allow moderate (SRP sequence and M13) or low (Sec sequence and M13) display efficiencies would select binders with higher binding affinities.
Protein directed evolution based on random mutagenesis and selection offers an alternative strategy to engineer Rubisco.
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