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We have also developed tools for the construction of conditional mutants where the expression of the target gene is regulated by an inducible promoter.
Here we show that a 20-amino acid peptide corresponding to Bax C-terminus (VTIFVAGVLTASLTIWKKMG) and two mutants where the two lysines are replaced with glutamate or leucine have potent membrane pore forming activities in zwitterionic and anionic phospholipid membranes.
Using a combination of VAP mutants where the cortical ER is detached from the PM and a set of artificial ER-PM tethers suppressing this phenotype, we demonstrate that the PM footprint of the cortical ER is functionally insulated from the cytosol.
In contrast, the level of Hcp was reduced in the ΔcqsA and ΔluxS mutants where the level of the hapR expression was expected to be reduced.
Importantly, the R cells are disrupted in this deficiency to an extent greater than the Dg point mutants where the R cells were simply stunted.
As such, a locus was considered to be beneficial or deleterious in aminoglycosides if i) the prevalence of mutants where the locus was disrupted changed significantly during the enrichments for at least two of the four drugs and ii) the z-scores for the locus for all four drugs had the same sign.
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Unlike xth31 mutant where the Al content of the whole roots is decreased nearly three folds, our ral1 mutant had reduced Al accumulation only in the root tips.
We have produced a mutant where the serine 35, a residue located at the entrance of the alcohol binding site, has been replaced by a cysteine residue.
Despite this, the Gap1-133 mutant where the LLKQ sequence has been mutagenized did not display increased sensitivity to D-His on Am medium.
Interestingly, the chitin synthesis related gene appears to be down-regulated in the ΔMovam7 mutant, in contrast to the ΔMosec22 mutant where the chitin synthesis related gene appeared to be up-regulated [18].
We next investigated bi-flagellated cells from the DNAI1RNAi mutant where the tip-to-base wave propagation is completely abrogated but that are still able to generate the base-to-tip wave.
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