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Panels show mutations detected in yeast transformants at the rDNA locus.
Because the specific gene(s) responsible for kanamycin resistance in T. neapolitana are unknown, we were unable to validate the possible T. sp.RQ7 transformants at the molecular level.
As expected from the growth pattern, specific growth rate of transformants at the initial period was significantly higher than the rest of the period.
In an effort to resolve this poor performance of transformants at the later stages of cultivation, we attempted a preliminary fed-batch experiment where nutrients were supplemented at intervals to one of the transformants NsbHLH2 3–11.
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A single-copy integration of each transformant at the PDR5 locus was confirmed by Southern hybridization (data not shown).
The growth rate of the MstG transformant during the exponential growth phase was comparable in the three different conditions, while in case of the MstH transformant, at the highest glucose concentration the growth rate was reduced.
They are a population of transformants at this point (i.e. not clonal) so we expect the proportion of individual insertions relative to one another may change over time.
Growth of the transformants at elevated glucose concentrations in the presence of fructose resulted in improved assimilation of the provided carbohydrates and a significant increase in the overall fermentation productivities.
However, none of the transformants produced progeny although some phenotypic variation was found with several transformants dying at the two leaf stage similar to the homozygous mutant and others developing up to the flowering stage.
The cells in which pJRSΔcsrS had been integrated into the chromosome were selected by the growth of the transformants at 39°C with erythromycin selection.
Under N limitation and osmotic stress, the DCW of the transformants at 8th day was more than 20%% greater than WT (Fig. 4a).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com