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In the complementation assay, we used the acrb-knockout, E. coli strain MG1655, referred to as Δacrb hereafter, as the background strain, which is sensitive to several antibiotics, including to ciprofloxacin.
A 6,334-bp 6,334-bpDNA fragenomicontaining the entire GhDNA coding region and upstream and downstream sequences ofragmentwas subcontainingm the BAC clonentire7658 and inserted into the binary vector pCAMBIA1300 to Ghd10ate the transformation vecodingGhd10 foregionin the complementandon test.
Plasmid pREP3Xdl was used as control in the complementation tests.
We also included the mutants T474A and T469A in the complementation assays to dissect the functional significance of these mutations in vivo.
This apparent gain of function did not extend to W121F, however, which was deficient in the complementation of both infection and yeast translation.
Expression of och1 in the complementation construct is driven by the constitutive gpdA promoter and is therefore not completely restoring the wild type situation.
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In contrast, the complementation hypothesis was supported in the children with autism, as well (children with deafness and SLI).
In addition, the complementation reaction with McLuc1 occurs within a few minutes and is reversible.
In summary, the complementation experiments confirm that Rpi-sto1 is a functional late blight R gene.
Here we describe, in detail, the complementation experiments for mutant 7K, as one example of the direct link between mutation and phenotype.
In order to further confirm the effect of ARP6 in AMT process, the complementation and overexpression assays were performed.
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