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The IE1 transactivator (hr3IE1) was included during transfection at a tenth of the total DNA amount to determine its potential to improve promoter activity or transposition frequency [ 29, 30].
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It was shown that integration of gene prediction and EST information improved promoter prediction [ 22, 38].
The methodology presented here should be valuable both to manipulate and improve yeast promoters, for promoter probe approaches, and for applied perspectives as well as to generate basic understanding of microbial promoters.
Further tuning may improve the promoter prediction slightly.
This emphasizes the need for effective methods to analyse and improve yeast promoter systems.
We have previously developed combinatorial mutagenesis and selection methods to improve bacterial promoter systems for diverse purposes.
However, it is often not feasible to transfect sufficient DNA required to achieve high expression levels or to improve the promoter to compensate for low expression.
To improve the promoter prediction, the use of DNA structural properties such as bendability, B-DNA twist, and duplex-free energy has been further explored for several eukaryotic genomes, including plants [ 10, 11].
CONCLUSION: The main finding, now supported by comprehensive data, is that the accuracy of human promoter predictors for high-throughput annotation purposes can be significantly improved if promoter prediction is combined with gene prediction.
FT activity improved with promoter addition in the order; K MoC1−x/Al2O3 > Na MoC1−x/Al2O3 > Ce MoC1−x/Al2O3 > Co MoC1−x/Al2O3 > MoC1−x/Al2O3 while chain growth probability varied with feed composition and was enhanced by all promoters.
In this work, the expression of CGTase from Geobacillus stearothermophilus in Escherichia coli BL21 (DE3) was significantly improved by promoter engineering and downstream box evolution.
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