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Because of extreme sensibility of E. coli to E protein [ 22], the normal growth of these strains indicated the tight control of PfhuA1 activity in different copy-numbered vectors under the repression condition.
Trying to find the key repressor of cellulase genes, we reasoned that such a regulator might be up-regulated under ER stress (mild as well as acute) and down-regulated under standard cellulolytic induction (Avicel only) compared to the repression condition on sucrose.
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Each of the 116 strains was submitted to growth assays (streaks and drops of serial dilutions on plate) under induction or repression conditions of bax and ORFs expression.
After two days on a galactose medium (induction conditions), differences in the growth of strains containing wild-type and mutated PrD were clearly confirmed for five mutants whereas they grew similarly on the glucose medium (repression conditions).
A single copy of the Cre gene ensures an extremely low leakage level of Cre recombinase expression under repression conditions.
Mechanistically, it is known that, under glucose repression conditions, acetic acid enters the cells by diffusion and at an acidic pH leads to intracellular acidification, anion accumulation and inhibition of cellular metabolic activity, namely fermentation and growth [ 2, 6].
Six half litre cultures of the recA- strain HB101Cre, six half litre cultures of the control recA+ strain MM219Cre and one half litre culture of the control recA+ strain TB1Cre, all of them harbouring the minicircle producer plasmid pFIXluc, were grown under catabolite repression conditions.
To further compare the gene expression changes between the Oct3/4-repression condition (ZHBTc4) and Oct3/4-overexpression condition (ZHTc6), we plotted the fraction of genes in each cluster of ZHTc6 cells present in each cluster of ZHBTc4 cells (Figure 1D).
This indicates that genes downregulated under Oct3/4-repression conditions tend to be also downregulated under Oct3/4-overexpression conditions.
Thus, genes upregulated under the Oct3/4-repression conditions tend to be also upregulated under the Oct3/4-overexpression conditions.
Gambia is set to hold an election this December, although due to political repression, voting conditions in the country are far from fair.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com