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We subsequently verified the phenotype-genotype relation by overexpression, reintroducing and complementing the mutation.
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To confirm the mutant data we included strains complementing the mutations with plasmids carrying the CgYAP1 and CgSKN7 genes regulated by their own promoters.
To complement the mutation, NIH-3T3 cells were stably transfected with the M115 open reading frame (gL-3T3).
Éric Déziel (personal communication) confirmed the mvfR phenotype of PA7, and complemented the mutation by adding a plasmid containing the wild-type mvfR gene from PAO1.
Taken together, these data demonstrated that the genomic fragment fully complemented the mutation.
Likewise the Pch PRP8 His5-intein was able to complement the mutation in HIS3 in this genetic background.
To complement the mutation data, and for a better overview of the activation status of the PI3K pathway, we investigated a set of previously published GSs.
After genotyping for the presence of the stm-dgh6 mutation, we could not distinguish the WT, stm-dgh6 heterozygotes and stm-dgh6 homozygote plants visually, confirming that pSTM STM-Venus can fully compSTM STM-Venusation.
Thus, dimerization of a subunit containing the inactivating mutation with a functional subunit can complement the mutation; this must occur either by lowering or by bypassing kinetic barriers in the ATPase or mechanical cycles of the mutant head.
These results indicate that VvZIP3 complemented the mutations of ZHY3, apparently by transporting Zn across the yeast plasma membrane.
The morphological effect of the irx mutations led us to hypothesize that the main reason for dwarfed growth in irx mutants is the collapsed vessels, and that specifically restoring xylan biosynthesis in vessels would therefore complement the mutations.
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