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An additional driver gene mutation increases a cell's fitness by a factor of (1 +s d ), whereas an additional housekeeper gene mutation decreases fitness by a factor of (1 −s h ) and the effect of multiple mutations is multiplicative.
Inactivation of the p53 pathway through multiple mutations is an almost universal feature of human cancer cells [ 50, 51].
The incidence of multiple mutations is significantly higher in heavy smokers than in non-smokers (P = 0.045).
For parE gene, the melting temperature for wild-type allele is at 84.6 84.9°C, whereas that of the mutants containing multiple mutations is at 84.9 85.2°C.
The potential effect of genetic variants should not be underestimated even if detected in unaffected individuals, because the combination of multiple mutations is probably the critical factor in human NTDs.
But as more random mutations are added, high-fitness genotypes become less common, and the average harm of multiple mutations is stronger than what the effect of single mutations indicated.
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How is change possible where multiple mutations are necessary but intermediate steps have no apparent advantage?
For example, in cavefish, we do not know what is the precise mutation that causes a change in gene expression level, or if multiple mutations are involved.
In this study, the bacterial laccase gene (cueO) with single or multiple mutations was directly cloned based on the metagenome of a chemical plant sludge.
However, multiple mutations are often deleterious for the activity and the identification of all the residues that historically led to the function is complicated.
Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins.
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