Genotyping of human alcohol dehydrogenases at the ADH2 and ADH3 loci following DNA sequence amplification

Humans are polymorphic at two of the alcohol dehydrogenase (ADH) loci important in ethanol metabolism, ADH2 and ADH3. Although the coding regions of these genes are 94% identical, they produce subunits that differ greatly in kinetic properties in vitro. These differences are likely to be reflected in the pharmacokinetics of alcohol metabolism, but studies have been hampered by the need to use liver biopsy specimens to determine the ADH phenotype. This problem has now been overcome by determining the genotype at these loci using DNA that has been amplified in vitro by the polymerase chain reaction. We report here the identification of all three of the ADH2 alleles and both of the ADH3 alleles. Any pair of ADH2 or ADH3 alleles can be distinguished using allele-specific oligonucleotide probes directed at their single base pair difference. In addition, ADH2(2) can be distinguished from ADH2(1) and ADH2(3) by detecting a new MaeIII site created in the third exon by the single base pair alteration in ADH2(2).     

Linkage disequilibrium and recent selection at three immunity receptor loci in Drosophila simulans

Immune system genes in a California population sample of Drosophila simulans were shown to bear several hallmarks of the effects of past directional selection. One potential effect of directional selection is an increase in linkage disequilibrium among the polymorphic sites that are linked to the site under selection. In this study, we focus on three D. simulans immunity loci, Hmu, Sr-CI/Sr-CIII, and Tehao, for which the polymorphic sites are in nearly perfect linkage disequilibrium, an unusual finding even with respect to other immunity genes sampled from the same lines. The most likely explanation for this finding is that, at each locus, two divergent alleles have been selected to intermediate frequencies in the recent past. The extent to which the linkage disequilibrium extends to the flanks of each of the immunity genes is minimal, suggesting that the favored mutations actually occurred within the immunity genes themselves. Furthermore, the excess linkage disequilibrium found in the California population is not found in an African D. simulans population sample and may be a result of novel pathogen-mediated selection pressures encountered during establishment of non-African populations.     

Linkage disequilibrium mapping of quantitative trait loci under truncation selection

As a dense map of single nucleotide polymorphism (SNP) markers are available, population-based linkage disequilibrium (LD) mapping or association study is becoming one of the major tools for identifying quantitative trait loci (QTL) and for fine gene mapping. However, in many cases, LD between the marker and trait locus is not very strong. Approaches that maximize the potential of detecting LD will be essential for the success of LD mapping of QTL. In this paper, we propose two strategies for increasing the probability of detecting LD: (1) phenotypic selection and (2) haplotype LD mapping. To provide the foundations for LD mapping of QTL under selection, we develop analytic tools for assessing the impact of phenotypic selection on allele and haplotype frequencies, and LD under three trait models: single trait locus, two unlinked trait loci, and two linked trait loci with or without epistasis. In addition to a traditional chi(2) test, which compares the difference in allele or haplotype frequencies in the selected sample and population sample, we present multiple regression methods for LD mapping of QTL, and investigate which methods are effective in employing phenotypic selection for QTL mapping. We also develop a statistical framework for investigating and comparing the power of the single marker and multilocus haplotype test for LD mapping of QTL. Finally, the proposed methods are applied to mapping QTL influencing variation in systolic blood pressure in an isolated Chinese population.     

Linkage disequilibrium analysis for microsatellite loci in six cattle breeds

Autosomal microsatellites are valuable tools for investigating genetic diversity and population structure and making conservation decisions to preserve valuable breeds of domestic animals. We carried out a linkage disequilibrium analysis using 29 microsatellite markers in six cattle populations: Suksun, Istoben, Yaroslavl, Kholmogory, Grey Ukrainian and Pechora type of Khologory cattle. We discovered a significant linkage disequilibrium between microsatellites INRA037 and CSRM60 in Grey Ukrainian cattle.     

Linkage disequilibrium between two genetic loci of the major histocompatibility complex in rats

The major histocompatibility complex (MHC) in natural populations of rats is composed of genetic phenotypes that are similar, if not identical, to those seen in inbred laboratory strains. Examination of individual wild rats from a single location in the city of Pittsburgh, Pennsylvania, resulted in the identification of seven different RT1.A histocompatibility serotypes and three RT1.B mixed lymphocytes responses. In this population of animals there is a significant association (p < 0.005) between four RT1.A and RT1.B phenotypic pairs: RT1.A⁸B¹, RTl.A^kBⁿ, RTl.A^dB^a and RT1.A¹B^a. The observed values for linkage disequilibrium (0.211,0.076,0.070 and 0.085, respectively) are very high and are close to the maximum expected, given the individual allelic frequencies. Although the animals included in this study were obtained from one location, agreement with Hardy-Weinberg equilibrium is demonstrated for other loci in the same population. The demonstration of equilibrium suggests that significant inbreeding is not affecting this population of rats. Not enough is known about the allelic frequencies in surrounding rat populations to determine how important the effect of migration is on these disequilibrium values. The large linkage disequilibria may indicate that, in the rat, environmental selective forces are operating to ensure the nonrandom association of separate components of the MHC.     

Linkage disequilibrium and modification of risk for Huntington disease.

The major limitation in performing predictive testing for Huntington disease (HD) is the unavailability of DNA from crucial family members. In our program approximately 20% (36/183) of persons have been excluded from predictive testing because of this reason. The major aim of this study was to examine whether data derived from linkage disequilibrium could modify risk analysis for persons at risk for HD. As a first step, we assessed whether the previously reported linkage disequilibrium between alleles recognized by probe pBS674E-D at locus D4S95 remained significant in a much larger data set. A total of 1,150 chromosomes from 622 individuals--200 affected and 422 unaffected--from 118 families were assessed. Significant haplotype association was detected with AccI and MboI RFLPs at the locus D4S95, with all the families (P = .00003), as well as for a subset from the United Kingdom (P = .0037). Data derived from linkage disequilibrium studies using D4S95 modifies the risk for HD, especially in persons of U.K. descent. Utilization of this approach for risk modification of HD awaits both validation of these data and additional information concerning ethnic-specific alleles at the D4S95 locus.     

Linkage disequilibrium fine mapping of quantitative trait loci: A simulation study

Recently, the use of linkage disequilibrium (LD) to locate genes which affect quantitative traits (QTL) has received an increasing interest, but the plausibility of fine mapping using linkage disequilibrium techniques for QTL has not been well studied. The main objectives of this work were to (1) measure the extent and pattern of LD between a putative QTL and nearby markers in finite populations and (2) investigate the usefulness of LD in fine mapping QTL in simulated populations using a dense map of multiallelic or biallelic marker loci. The test of association between a marker and QTL and the power of the test were calculated based on single-marker regression analysis. The results show the presence of substantial linkage disequilibrium with closely linked marker loci after 100 to 200 generations of random mating. Although the power to test the association with a frequent QTL of large effect was satisfactory, the power was low for the QTL with a small effect and/or low frequency. More powerful, multi-locus methods may be required to map low frequent QTL with small genetic effects, as well as combining both linkage and linkage disequilibrium information. The results also showed that multiallelic markers are more useful than biallelic markers to detect linkage disequilibrium and association at an equal distance.     

Linkage disequilibrium, selection and recombination at three Loci

Limits to the relationship among linkage disequilibrium, selection and recombination at equilibrium in three-locus, two-allele, deterministic, discrete generation models are determined using linear programming techniques. These results show that the commonly used measures of linkage disequilibrium are not appropriate for a multilocus setting. Additionally, interactions among three loci are important in reducing the strength of selection necessary to maintain a given level of disequilibrium, relative to a two-locus model.     

Linkage disequilibrium at the Angelman syndrome gene UBE3A in autism families.

Autistic disorder is a neurodevelopmental disorder with a complex genetic etiology. Observations of maternal duplications affecting chromosome 15q11-q13 in patients with autism and evidence for linkage and linkage disequilibrium to markers in this region in chromosomally normal autism families indicate the existence of a susceptibility locus. We have screened the families of the Collaborative Linkage Study of Autism for several markers spanning a candidate region covering approximately 2 Mb and including the Angelman syndrome gene (UBE3A) and a cluster of gamma-aminobutyric acid (GABA(A)) receptor subunit genes (GABRB3, GABRA5, and GABRG3). We found significant evidence for linkage disequilibrium at marker D15S122, located at the 5' end of UBE3A. This is the first report, to our knowledge, of linkage disequilibrium at UBE3A in autism families. Characterization of null alleles detected at D15S822 in the course of genetic studies of this region showed a small (approximately 5-kb) genomic deletion, which was present at somewhat higher frequencies in autism families than in controls.     

Linkage disequilibrium and gametophytic self-incompatibility

Summary The approach to linkage equilibrium of a locus linked to the locus determining gametophytic self-incompatibility (S) is considered. For the simplest case of three alleles at the S locus and two at the linked locus it is necessary to consider 3 measures of linkage disequilibrium. These are found to approach their equilibrium value of zero in one of three ways: 1) steadily declining to zero; 2) oscillating as decline proceeds; 3) a combination: 2) followed by 1). Linkage equilibrium may be established before genotype frequencies reach their expectation under random crossing. Earlier studies (Li 1951; Moran 1962) of the approach to S allele equilibrium have been based on the assumption that all types of pollen take part in fertilizations equally frequently. Such an assumption leads to simpler expressions for changes in S gene frequencies but is extremely unrealistic and, in particular, leads to a different rate of approach to equilibrium from the more comprehensive model. It is shown that even in the absence of selection it is not possible to predict the equilibrium gene frequency of a linked locus until S allele equilibrium is reached. This frequency may be either higher or lower than that calculated from a gene count in the starting genotype pool. However, these two gene frequencies may stabilize long before linkage equilibrium is achieved. An examination of selection against one genotype at the linked locus is undertaken. If linkage is complete, lethality can be less effective at reducing the gene frequency than is less intense selection (in only a few generations of selection). Here too linkage equilibrium may be established with selection still effective in bringing about a decline in gene frequency. An examination of the analysis and conclusions of Rasmuson (1980) shows that because these were based on the inadequate formulae previously discussed and exclude phenomena discussed above, they are misleading. The possibility of a gametophytic self-incompatibility system providing a sufficient condition for the sheltering of lethals in the absence of the condition of complete linkage to the S locus (r=0) is shown to be unlikely.     

Two loci with two alleles: Linkage equilibrium and linkage disequilibrium can be simultaneously stable

A class of two-locus two-allele viability matrices is exhibited which has the property that, for a large range of recombination values, both linkage equilibrium and linkage disequilibrium are stable. The model is specified by five viabilities; the classical schemes previously analyzed involve at most four selection parameters.     

Linkage disequilibrium with the island model

Linkage disequilibrium between two linked loci was studied for a finite population with a subdivided population structure. Wright's island model was used; extinction and replacement of colonies were also incorporated. Two alleles (A₁ and A₂ at the first locus, and B₁ and B₂ at the second locus) with symmetric mutation rates were assumed, and equilibrium properties of linkage disequilibrium coefficients were analyzed. In terms of analogy with the subdivision of inbreeding coefficient, the variance of linkage disequilibrium is divided into several components: D²_IS (variance of within-colony disequilibrium), D²_ST (variance of correlation of A₁ and B₁ of different gametes of one colony relative to that of the total population), and D²_IT (total variance of disequilibrium). Other subdivisions are D'²_IS (variance of correlation of A₁ and B₁ of one gamete of a colony relative to that of the average gamete of the population) and D'²_ST (variance of the ordinary disequilibrium of the whole population). When migration is limited, the variance becomes large if the correlation of A₁ and B₁ of one colony is taken relative to that of the whole population (D²_ST and D'²_IS). Also, when the rate of extinction-replacement of colonies is high, the whole-population disequilibrium coefficient (D'²_ST) can become fairly large. Observed linkage disequilibria, such as those among markers in the major histocompatibility complex of man and mouse, may well be explained by limited migration, without assuming epistatic natural selection.     

Linkage disequilibrium in the domesticated pig

This study investigated the extent of linkage disequilibrium (LD) in two genomic regions (on chromosomes 4 and 7) in five populations of domesticated pigs. LD was measured with D' and tested for significance with the Fisher exact test. Effects of genetic (linkage) distance, chromosome, population, and their interactions on D' were tested both through a linear model analysis of covariance and by a theoretical nonlinear model. The overall result was that (1) the distance explained most of the variability of D', (2) the effect of chromosome was significant, and (3) the effect of population was significant. The significance of the chromosome effect may have resulted from selection and the significance of the population effect illustrates the effects of population structures and effective population sizes on LD. These results suggest that mapping methods based on LD may be valuable even with only moderately dense marker spacing in pigs.     

Linkage disequilibrium between alleles at highly polymorphic mini- and micro-satellite loci of Theileria parva isolated from cattle in three regions of Kenya

Theileria parva schizont-infected lymphocyte culture isolates from western, central and coastal Kenya were analysed for size polymorphism at 30 T. parva-specific variable number tandem repeat (VNTR) loci using a panel of mini- and micro-satellite markers. The mean number of alleles ranged from 3 to 11 at individual loci and 183 distinct alleles were observed in total, indicating high genetic diversity within the T. parva gene pool in Kenyan cattle. The frequency distribution of the length variation of specific alleles among isolates ranged from normal to markedly discontinuous. Genetic relationships between isolates were analysed using standard indices of genetic distance. Genetic distances and dendrograms derived from these using neighbour-joining algorithms did not indicate significant clustering on a geographical basis. Analysis of molecular variance demonstrated that the genetic variation between individual isolates was 72%, but only 2.3% when isolates from different regions were pooled. Both these observations suggest minimal genetic sub-structuring relative to geographical origin. Linkage disequilibrium was observed between pairs of loci within populations, as in certain Ugandan T. parva populations. A novel observation was that disequilibrium was also detected between alleles at three individual pairs of VNTR loci when isolates from the three regional meta-populations were pooled for analysis.