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Dr Drayna's analysis suggests that stuttering is linked to mutation of a gene called GNPTAB a finding confirmed when the same mutation was subsequently discovered in a number of stutterers from Pakistan and India who were unrelated to the original group of families.
The mutation was subsequently confirmed by conventional Sanger sequencing in fibroblast-derived germline DNA; thus, the diagnosis of Li-Fraumeni syndrome (LFS, OMIM #151623) was made.
The resulting nuc2 mutation was subsequently introduced into the isogenic nuc1, sarA and sarA/nuc1 mutants by phage-mediated transduction [7].
Pseudogenes contained one or more mutations that would ablate expression; each inactivating mutation was subsequently checked against the original sequencing data.
The R46X transcript mutation was subsequently identified in several unaffected subjects in variable fractions by direct sequencing of PBMC cDNAs (Fig. 1B).
Each intron mutation was subsequently confirmed by sequencing.
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The DNA fragments between −450 and −206 of RETN with or without this point mutation were subsequently cloned into a luciferase reporter vector pGL4.11 (Promega) at the site between Kpn I and Bgl II.
The resulting H3 variant as well as a control protein synthesized with unmodified Ser10 and also containing the T32C mutation were subsequently incorporated into nucleosome arrays.
Pseudogenes had one or more mutations that would prevent correct translation; each of the inactivating mutations was subsequently checked against the original sequencing data.
The presence of slightly deleterious mutations was subsequently identified as the likely reason for the excess of nonsynonymous polymorphism, compared with divergence, in mitochondrial protein-coding genes (Nachman 1998; Rand and Kann 1998; Gerber et al. 2001).
Combinatorial mutations were subsequently introduced, and the superior mutant was double mutant E161D/N365P.
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