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Although Arabidopsis mutants that represent multiple mutation alleles of AtPP2 have been generated [ 27, 28], subsequent biological effects have not been studied, and especially, effects of AtPP2 mutations on the plant resistance to insects are unclear.
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Other studies have used strains with extremely high mutation rates due to multiple mutator alleles (Larrea et al. 2010).
In agreement with the presence of up to 7 gene copies and the subsequent need for multiple mutated alleles to visualize an effect on macrolide resistance9, in the present study no mutations in the 23S rRNA gene were observed in the 66 isolates analysed.
A total of 71 mutations were identified by TILLING, providing multiple mutant alleles for every gene tested.
A region of approximately 1500 bp from each gene was examined and a total of 71 novel mutations were identified by TILLING, thus providing multiple mutant alleles for each gene (Table 1).
This can increase the chance of unwanted off-target effects as well as on-target mutations and mutational mosaicism in independent cells of embryos, leading to multiple mutant alleles in following generations and potentially complicating further analyses.
Several SNPs located in 5% MCS regions were eliminated from the study due to the nature of the variation precluding genotyping on the Illumina platform (e.g. insertion/deletions or multiple mutation events leading to > 2 alleles) and/or the failure to generate an Illumina score > 0.60.
This is even less probable when the private alleles/haplotypes diverge by more than one mutation from other alleles/haplotypes, since this would involve multiple mutation events.
If alleles carry multiple mutations, you may need to perform site-directed mutagenesis to create single amino acid substitution alleles and test their ability to confer drug resistance.
Multiple mutations are necessary for carcinogenesis, and as APC mutations occur in a large majority of human colorectal cancers, FBXW7 mutations are found mostly in the context of APC mutant alleles.
Are multiple mutations required?
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