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The recircularized plasmid is then rescued from yeast transformants that show robust protein expression in the yeast cultures on the same day.
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The only transformants that showed the large open vessels characteristic for the wild type were the ones expressing the pVND7 IRX7 construct.
The use of rice chi11, encoding a chitinase, as a co-transgene along with tlp produced a tlp chi11 co-transformant that showed enhanced resistance against R. solani than the ones that express tlp or chi11 transgene alone.
The only transformant that showed a significant growth rate (with an approximate doubling time of 3 h) was the one encoding the K617A mutant (Fig. 4).
However, the relatively low number of bands in each restriction digest of genomic DNA from the two transformants that are shown in Figure 7D and from four other transformants that were analyzed, suggests that there are only ~1 3 integrations of the co-transformed plasmid, so a high gene dosage through a large number of integrations can be excluded.
Our results show transformants that overexpress the modified pfkA gene in combination with cadA have significantly increased citrate secretion levels.
Out of 192 transformants that were screened, four showed a single 2490 bp amplicon, which confirmed that the deletion construct had integrated into the KU80 locus (Fig. S3B).
Transformants were obtained that showed significantly higher secreted protein yield, with a resulting heterologous protein activity that ranged between 10% and 373% higher compared to the parental strains when grown in complex media.
Transformants that were generated through co-transformation with plasmid pHsp70A-GLuc were also analyzed qualitatively for luciferase activity, and several of these transformants showed chemiluminescence (data not shown).
In the present study, different IBE-producing transformants of C. acetobutylicum that showed high isopropanol excretion capacities have been constructed.
The molecular analysis of transformants showed that the integration was ectopic (data not shown), supporting the hypothesis that SesB alone is per se a functional gene involved in Secteur development.
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