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Overexpression of either RHEBp.P37L or p.S68P, also resulted in significantly increased headsize, reminiscent of the megalencephaly seen in our patients (p < 0.0001 for either mutant allele when compared to WT RHEB; Fig. 3d, e).
Thus, our working hypothesis is that altered biophysical properties of double mutant sarcomeric proteins could interact to either heighten or mitigate the functional cellular outcomes of either mutant allele when studied in isolation.
We could detect a 20-bp deletion mutation in a pool of DNA containing as little as 5% of a mutant allele when we used T7E1.
A typical formalin-fixed paraffin-embedded colorectal cancer specimen might have between 2-50% tumor cells, which corresponds to 1-25% mutant allele when the tumor is diploid and a heterozygous mutation is present.
However, we found a much reduced transmission frequency of the cop1-5 mutallelelele when compared to the COP1 wild-type allele, thus making the analysis of segregating populations ambiguous.
DOI: http://dx.doi.org/10.7554/eLife.04875.008 We performed three tests to investigate the role of the different mutations in producing the evolved phenotype: deleting the mutated gene, replacing it with a wild-type allele, and investigating selection for or against the mutant allele when we selected for different levels of cell association.
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The consistency of mutant allele ratios, when testing different random subareas, would further support that concomitant mutations are present in the same population.
The ES cells and mice that we have created carry a silenced RIα mutant allele (RIαB) that, when activated by Cre recombinase, expresses an RIα protein that is defective in cAMP binding.
Wings from another dSno mutant allele dSnoGS-c517 [10] when in trans to dSnoEx4B also exhibit ectopic margin bristles in the anterior compartment and ectopic sensilla (n = 136; Figure S4F).
Microscopic examination revealed that worms carrying either mutant hcf-1 allele when grown at 12°C frequently display cells with multiple nuclei of heterogeneous size, extra spindle poles and improper spindle alignment; the embryos often stopped development at or prior to gastrulation (data not shown).
The parameter H captures the degree of dominance of the mutant allele, which is fully recessive when H = 0, co-dominant when H = 1/2, and fully dominant when H = 1 [ 13].
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