Analysis of gene complexes predisposing to coronary atherosclerosis

The following seven polymorphic marker loci of genes responsible for predisposition to coronary atherosclerosis (CAS) were studied: the ACE locus responsible for angiotensin-converting enzyme insertion/deletion polymorphism for the presence or absence of the Alu insertion in the gene; the F13, PLAT, and APOA1 loci, controlling the clotting factor 13, plasminogen-activating tissue factor, and apolipoprotein A, respectively; the MTHFR and AGT polymorphic loci responsible for point mutations in methylenetetrahydrofolate reductase and those in angiotensinogen, respectively, and the NOS3 locus controlling the number of tandem repeats in the nitric oxide synthase gene. These loci are located on different chromosomes and encode products involved into various metabolic pathways leading to CAS. In the populations studied, significant differences between healthy subjects and patients predisposed to cardiovascular diseases were revealed with regard to the above seven markers. The 174M allele (T174M polymorphism in the ACE gene) was significantly associated with coronary atherosclerosis. It was found that specific gene combinations are involved in the CAS development and determine variation in the pathogenetically important quantitative traits.     

Isolation of polymorphic microsatellites in the stemless thistle (Cirsium acaule) and their utility in other Cirsium species

The genus Cirsium includes species with both widespread and restricted geographical distributions, several of which are serious weeds. Nine polymorphic microsatellite loci were isolated from the stemless thistle Cirsium acaule. Eight were polymorphic in C. acaule, six in C. arvense and seven in C. heterophyllum. One locus monomorphic in C. acaule showed polymorphism in C. heterophyllum. The mean number of alleles per locus was 4.1 in C. acaule, 6.2 in C. arvense and 2.9 in C. heterophyllum. These nine loci were also amplified in C. eriophorum and C. vulgare, suggesting that these markers may be of use throughout the genus.     

Development of microsatellite markers in the toxic dinoflagellate Alexandrium tamarense (Dinophyceae)

Outbreaks of paralytic shellfish poisoning caused by the toxic dinoflagellate Alexandrium tamarense (Dinophyceae) are currently a serious problem from an economic and food hygiene point of view throughout the world. We isolated 13 polymorphic microsatellite loci from this species. These loci provided microsatellite markers with high polymorphism ranging from four to 15 alleles per locus and gene diversity between 0.632 and 0.974. The markers are available for more detailed investigations of genetic structure and gene flow of A. tamarense populations.     

Connection of the polymorphic variant T174M angiotensinogen gene with coronary atherosclerosis in the Tomsk population

Allele and genotype frequencies of the T174M polymorphism of the angiotensinogen gene were for the first time estimated in the West Siberian population. The polymorphism was tested for association with coronary atherosclerosis (CAS) and with several quantitative risk factors in patients with angiographically verified CAS, healthy individuals, and in a population sample nondifferentiated with respect to CAS.     

Isolation and characterization of polymorphic microsatellite loci in the gudgeon,Gobio gobio (Cyprinidae)

A total of seven polymorphic microsatellite loci from Gobio gobio were isolated and characterized. A preliminary population survey of 82 specimens from four populations, located in a downstream pollution gradient of cadmium and zinc, demonstrated the usefulness of these primers both in population genetic studies in general, as well as in evaluating the effects of anthropogenic pollution on genetic structure. Overall locus polymorphism ranged from two to 13 alleles. Observed heterozygosity per locus ranged from 0.39 to 0.73.     

Association between Polymorphism T174M of the Angiotensinogen Gene and Coronary Atherosclerosis in the Tomsk Population

Allele and genotype frequencies of the T174M polymorphism of the angiotensinogen gene were for the first time estimated in the West Siberian population. The polymorphism was tested for association with coronary atherosclerosis (CAS) and with several quantitative risk factors in patients with angiographically verified CAS, healthy individuals, and in a population sample nondifferentiated with respect to CAS.     

Gene polymorphisms for elucidating the genetic structure of the heavy-metal hyperaccumulating trait in Thlaspi caerulescens and their cross-genera amplification in Brassicaceae

Genetic polymorphism was investigated in Thlaspi caerulescens J. & C. Presl at 15 gene regions, of which seven have been identified to putatively play a role in heavy-metal tolerance or hyperaccumulation. Single nucleotide and length polymorphisms were assessed at four cleaved amplified polymorphic sequences (CAPS) and 11 simple sequence repeat (microsatellite) loci, respectively. The utility of these loci for genetic studies in T. caerulescens was measured among seven natural populations (135 individuals). Fourteen loci rendered polymorphism, and the number of alleles per locus varied from 2 to 5 and 1 to 27 for CAPS and microsatellites, respectively. Up to 12 alleles per locus were detected in a population. The global observed heterozygosity per population varied between 0.01 and 0.31. Additionally, cross-species/genera amplification of loci was investigated on eight other Brassicaceae (five individuals per population). Overall, 70% of the cross-species/genera amplifications were successful, and among them, more than 40% provided intraspecific polymorphisms within a single population. This indicates that such markers may, as well, allow comparative population genetic or mapping studies between and within several Brassicaceae, particularly for genes involved in traits such as heavy-metal tolerance and/or hyperaccumulation.     

Dinucleotide microsatellite markers from Anopheles funestus

A total of 39 dinucleotide microsatellite‐containing clones were sequenced from a principal African malaria vector, Anopheles funestus. Primers designed to amplify 20 loci were used to genotype A. funestus mosquitoes from Burkina Faso and Kenya. Of nine polymorphic loci that amplified reliably and could be scored unambiguously, the overall within‐sample gene diversity was similar between locales, 0.77 and 0.78, with an allelic richness per locus of five to 11. Both the high level of polymorphism and absence of significant heterozygote deficiency at any locus favour these markers for studies of population structure that are vital to controlling this medically important species.     

Genomics of Type 1 Diabetes Mellitus and Its Late Complications

In ethnic Russians, MHC (HLA) was shown to be the major locus determining the predisposition to type 1 diabetes mellitus (T1DM). To map the regions linked to T1DM, families with concordant or discordant sib pairs were selected from the Russian population of Moscow. With these families, linkage to T1DM was demonstrated for the CTLA4 gene (IDDM12, 2q32.1–q33), which codes for a T-cell surface antigen, and the PDCD2 gene (IDDM8, 6q25–q27), which is homologous to the mouse programmed cell death activator gene. Using polymorphic microsatellites, we could also observe the linkage to T1DM of regions 3q21–q25 (IDDM9) and 10p12.2 (IDDM10). Complex analysis of linkage and association of the polymorphic markers from region 11p13 in the vicinity of the catalase gene (CAT) based on the case/control groups and two groups of families allowed us to reveal a new T1DM locus; the linkage to this locus was not reported earlier for other populations. Diabetic polyneuropathy (DPN) proved to be associated with polymorphic markers Ala(–9)Val of the SOD2 gene, Arg213Gly of the SOD3 gene, and T(–262)C of the CAT gene, and with a polymorphic microsatellite located in the promoter region of the NOS2 gene. It has been supposed that one of the main risk factors of DPN development in patients with type 1 diabetes is oxidative stress arising in hyperglycemia because of increased production of superoxide radicals in mitochondria and insufficient activity of antioxidative enzymes. Diabetic nephropathy (DN) showed no association with the antioxidative enzyme genes. However, the association was observed for the insertion/deletion (I/D) polymorphism of ACE and the ecNOS34a/4b polymorphism of NOS3, two genes involved in controlling vascular tonus, as well as for the I/D polymorphism of APOB and the 2/3/4 polymorphism of APOE, two genes involved in lipid transport. In addition, polymorphic microsatellites of chromosome 3q21–q25 proved to be closely associated with DN. The tightest association was established for D3S1550, carriers of allele 12 or genotype 12/14 having high risk of DN (OR = 4.85 and 6.25, respectively). Region 3q21–q25 perhaps contains a major gene determining DN development, while the other DN-associated genes mostly influence the progression of DN.     

Polymorphic microsatellite markers for chromosomal races of Australian morabine grasshoppers (Vandiemenella,viatica species group)

Chromosomally diverse Australian morabine grasshoppers (genus Vandiemenella, viatica species group) have parapatric distributions and occasionally hybridize at contact zones. To investigate population genetic structure and the extent of gene flow between chromosomal races/species of Vandiemenella, we isolated and characterized nine polymorphic microsatellite loci and one insertion/deletion polymorphic locus. The numbers of alleles per locus ranged from two to 34 across three chromosomal races on Kangaroo Island, South Australia, and expected heterozygosity within races ranged from 0.00 to 0.94. Inter‐taxon amplification was generally successful within Vandiemenella, but not for other morabine genera.     

Development and characterization of 13 novel microsatellite loci from the flat-headed bat (Tylonycteris pachypus) with cross-species amplification in closely related taxa

13 novel microsatellite loci was isolated using the enriched library method from genomic DNA of the flat-headed bat (Tylonycteris pachypus). These loci showed high levels of genetic polymorphism testing on 54 individuals sampled from Guangxi province, China. The number of observed alleles per locus ranged from 2 to 15. The expected and observed heterozygosity values ranged from 0.170 to 0.900 and from 0.185 to 0.944, respectively. One loci departed significantly from Hardy-Weinberg equilibrium (HWE) after Bonferroni correction and no Linkage disequilibrium was found between any pairs of loci. In addition, these loci were tested in the sister species, Tylonycteris robustula, seven loci amplified successfully and were also polymorphic.     

Development of microsatellite markers in the Japanese pear (Pyrus pyrifolia Nakai)

Thirteen polymorphic microsatellite loci were developed in the Japanese pear (Pyrus pyrifolia Nakai) by using an enriched genomic library. The obtained microsatellite loci showed a high degree of polymorphism in the Japanese pear with 3–6 alleles per locus. The average values of observed and expected heterozygosities among these 13 loci were 0.69 and 0.71, respectively. Ten microsatellites could successfully amplify loci in the European pear (Pyrus communis L.), which were highly polymorphic as well.     

Microsatellite loci for the Australian field cricket Teleogryllus oceanicus and their cross‐utility in Teleogryllus commodus

Microsatellite loci were isolated from the Australian field cricket Teleogryllus oceanicus. Seven polymorphic loci were found with an observed number of alleles ranging from eight to 17 and observed heterozygosities between 0.26 and 0.94. One locus was found to be X‐linked. These seven loci were also tested for amplification and polymorphism in the congeneric species Teleogryllus commodus. The loci will be used for paternity studies in these species.     

Polymorphic and monomorphic serum esterase heterogeneity in catostomid fish populations

A one-locus, two-allele serum esterase polymorphism is described in populations of Catostomus (Pantosteus) clarki in the connected and disconnected tributaries of the lower Colorado River system. A clinal distribution of allelic frequency is linearly correlated with latitude. This distribution of allelic frequencies, and other characteristics of the polymorphism in C. (P.) clarki populations, are indicative of the selective maintenance of the heterogeneity. Sympatric species of the subgenus Catostomus exhibit a monomorphic esterase phenotype indistinguishable from the heterozygote phenotype in polymorphic populations of C. (P.) clarki. Genetic mechanisms involved in the evolution of one polymorphic locus and/or two homozygous loci from an ancestral gene are discussed.     

Gene polymorphism of the renin-angiotensin system in six ethnogeographic regions of Belarus

The insertion/deletion polymorphism of the angiotensin-converting enzyme gene (ACE) and the T174M polymorphism of the angiotensinogen gene (AGT) have been studied in six ethnogeographic regions of Belarus. Significant intrapopulation differences in ACE genotype frequencies have been found for the northern and eastern regions (the Dvina and Dnepr basins, respectively). Significant differences in the AGT genotype frequencies have been found between populations of the Dnepr basin and populations of all other Belarusian regions. The allele and genotype frequencies of the genes studied in the Belarusian population and populations of other regions of the world have been compared. The frequencies of the insertion (I) and deletion (D) alleles of the ACE gene in the Belarusian population are 50.7 and 49.3%, respectively, which is similar to these frequencies in European countries. The frequency of the M allele of the AGT gene in Belarus is 16.6%, which is higher than its frequency in populations of European, African, and Asian origins.     

Rare McArdle disease locus polymorphic site on 11q13 contains CpG sequence

Summary When probes throughout the McArdle disease (myophosphorylase) gene region were used to search for DNA polymorphisms, only an MspI polymorphism was found in 94 enzyme-probe combinations. Along with an insertion/deletion polymorphism more 3 to the gene locus, these polymorphisms will be informative in 75% of at-risk patients. These results contrast strikingly to the six polymorphic sites detected in 15 enzyme-probe combinations in the homologous Her's disease (liver phosphorylase) gene region. This single MspI polymorphic site includes a CpG sequence of known increased mutability suggesting that DNA regions with rare polymorphisms will have most polymorphic sites at sequences with enhanced mutability. Fluorescence in situ hybridization sublocalized this gene to proximal band 11q13, establishing a point of cross-reference between the physical and genetic maps.     

Isolation of polymorphic microsatellite markers in the sub‐Saharan tree,Acacia (Senegalia) mellifera (Fabaceae: Mimosoideae)

We isolated 11 polymorphic microsatellite markers from Acacia mellifera, a savannah woodland tree in sub‐Saharan Africa and southern Arabia. The loci were screened for polymorphism using 48 Kenyan individuals. Allelic diversity ranged from three to 19 per locus and the polymorphic information content varied from 0.287 to 0.893. These loci will be useful in studies of genetic structure, gene flow and breeding systems.     

Isolation and characterization of polymorphic microsatellite loci for the study of paternity and population structure in the little penguin Eudyptula minor

We isolated and characterized eight novel microsatellite loci in the little penguin Eudyptula minor, using nonradioactive polymerase chain reaction‐based techniques to screen GA and GAAA repeats from enriched genomic DNA libraries. All eight loci were polymorphic and seven were variable in our main study population (mean H_E = 0.613, mean N_A = 7.14). Cross‐amplification using a microsatellite primer developed in Spheniscus demersus (African penguin) yielded one additional polymorphic locus. This locus combined with six of the little penguin loci is suitable for paternity assignment in little penguins (exclusion probability for seven unlinked loci = 0.993).     

Development of microsatellite markers in black locust (Robinia pseudoacacia) using a dual‐supression‐PCR technique

Black locust (Robinia pseudoacacia) is an economically and ecologically important tree species in the world. We isolated seven polymorphic microsatellite loci from R. pseudoacacia using a dual‐suppression‐PCR technique. These loci provided microsatellite markers with high polymorphism ranging from three to 12 alleles per locus and expected heterozygosity between 0.538 and 0.944. The markers are now available for more detailed investigation of population genetic structure and pollen and seed dispersal.