36 Although mutation is responsible for creating SNPs, their maintenance probably depends on natural selection on coding sequences, which in turn is regulated by its precise functional role as well as meiotic recombination. This marked variability of variability in candidate genes is also illustrated by the fact that extremely invariable gene regions can also occur, with no structural mutations at all, singletons, or complete absence of any variant in coding or regulatory regions, even when genes

were systematically resequenced in substantial numbers up to about 200 individuals.62,78,79 In particular, Inhibitors,research,lifescience,medical an extensive survey by Halushka et al36 showed that about, 17% of all genes were invariable at the protein level, which is in agreement with our extrapolations of a fraction of about 20% of invariable genes (Hoehe M et al, unpublished results). This may be related to certain aspects of yet, unknown or particularly high functional significance among the total gene pool,

and is one of the important questions to be addressed in the future. Taken together, Inhibitors,research,lifescience,medical there is no a priori way to predict the actual natural variability of a gene; it must be Bcl-2 inhibitor empirically assessed in appropriately Inhibitors,research,lifescience,medical chosen samples in each case. An example of the variability of variability in candidate genes, which may exist even within members of the same gene family (such as G protein-coupled receptor genes) or even within members of the same group of receptor subtypes or genes encoding endogenous receptor ligands, is given in Figure 3 80 These genes may represent prototype examples of drug targets and their potential Inhibitors,research,lifescience,medical variability, particularly with respect, to the fact that more than 50% of the total of 417 receptor targets of pharmaceutical relevance encode G protein-coupled receptors.4 Figure 3 Polymorphic spectra of candidate genes.80 The genomic reference sequences are presented as baseline, exonic sequences

as gray or white (untranslated regions [UTRs]) Inhibitors,research,lifescience,medical bars; sequences were drawn to scale, which is indicated. All gene variants are specified … For instance, in the human µ-opioid receptor gene (OPRM1) (Figure 3a), a target for morphine, the classical pain killer in contemporary medicine and substance of abuse, a total of 43 variants have been identified within 6.7 kb in 250 European- and African-Americans.29 Clearly, the 5′ regulatory and 5′UTR regions these (one SNP every 99 bp and 73 bp, respectively) and the critical regions in intron 2 (one SNP every 110 bp) were much more variable than the coding exon (one SNP every 267 bp) and other intronic regions. Five of the six SNPs in the coding region clearly affected the encoded protein; two of those (which were relatively frequent) were located in the N-terminal, one in the third transmembrane domain, and two in the third cytoplasmic loop; all were shown to induce functional alterations.