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Vol 52, No 10 (2016)

Molecular Genetics

Induction of transcription through the scs insulator leads to abnormal development of Drosophila melanogaster

Kyrchanova O.V., Leman D.V., Toshchakov S.V., Utkina M.V., Tikhonov M.V., Parshikov A.F., Maksimenko O.G., Georgiev P.G.

Abstract

A regulatory element named scs is one of the first insulators discovered in Drosophila, which was found on the boundary of the hsp70 domain. The 993-bp scs insulator contains two promoters at the ends and two polyadenylation signals located in the same orientation in the central part of the insulator. In the Drosophila transgenic lines, induction of a strong transcription through the scs insulator only in the direction that coincides with the direction of the two polyadenylation sites activity results in multiple phenotypic defects of the Drosophila development and embryonic lethality. A similar effect was not observed upon testing of other known Drosophila insulators.

Russian Journal of Genetics. 2016;52(10):1007-1014
pages 1007-1014 views

Genetics of Microorganisms

Occurrence of islands in genomes of Sinorhizobium meliloti native isolates

Muntyan V.S., Cherkasova M.E., Andronov E.E., Simarov B.V., Roumiantseva M.L.

Abstract

Genomes of 184 Sinorhizobium meliloti native isolates were studied to test the occurence of islands Sme21T, Sme19T, and Sme80S previously described in the model strain Rm1021. This analysis was conducted using PCR methodology involving specific primers. It was demonstrated that, in the examined geographically distinct populations of S. meliloti from the Northern Caucasus (NCG) and the Aral Sea region (PAG), the strains containing genomic islands were observed with similar frequency (0.55 and 0.57, respectively). Island Sme80S, denoted as an island of “environmental adaptivity,” was identified predominantly (frequency of 0.38) in genomes of strains which exhibited a lower level of salt tolerance and was isolated in PAG, a modern center of introgressive hybridization of alfalfa subjected to salinity. Island Sme21T designated as “ancestral” was observed in genomes of strains isolated in NCG, the primary center of host-plant biodiversity, 10-fold more often than in strains from PAG. An island Sme19T, which predominantly carries genes encoding transposases, was observed in genomes of strains in both populations with average frequency of 0.10. The analysis of linkage disequilibrium (LD) based on the assessment of probability for detection of different islands combinations in genomes revealed an independent inheritance of islands in salt-sensitive strains of various geographic origin. In contrast, the absence of this trend was noted in the majority of the examined combinations of salt-tolerant strains. It was concluded that the structure of chromosome in PAG strains which predominantly possessed a salt-sensitive phenotype was subjected to active recombinant processes, which could predetermine the intensity of microevolutionary processes in bacterial populations and facilitate an adaptation of bacteria in adverse environmental effect.

Russian Journal of Genetics. 2016;52(10):1015-1022
pages 1015-1022 views

Plant Genetics

Genetic polymorphism of Tulipa gesneriana L. evaluated on the basis of the ISSR marking data

Kashin A.S., Kritskaya T.A., Schanzer I.A.

Abstract

Using the method of ISSR analysis, the genetic diversity of 18 natural populations of Tulipa gesneriana L. from the north of the Lower Volga region was examined. The ten ISSR primers used in the study provided identification of 102 PCR fragments, of which 50 were polymorphic (49.0%). According to the proportion of polymorphic markers, two population groups were distinguished: (1) the populations in which the proportion of polymorphic markers ranged from 0.35 to 0.41; (2) the populations in which the proportion of polymorphic markers ranged from 0.64 to 0.85. UPGMA clustering analysis provided subdivision of the sample into two large clusters. The unrooted tree constructed using the Neighbor Joining algorithm had similar topology. The first cluster included slightly variable populations and the second cluster included highly variable populations. The AMOVA analysis showed statistically significant differences (FCT = 0.430; p = 0.000) between the two groups. Local populations are considerably genetically differentiated from each other (FST = 0.632) and have almost no links via modern gene flow, as evidenced by the results of the Mantel test (r =–0.118; p = 0.819). It is suggested that the degree of genetic similarities and differences between the populations depends on the time and the species dispersal patterns on these territories.

Russian Journal of Genetics. 2016;52(10):1023-1033
pages 1023-1033 views

Allelic diversity of hordein-coding loci Hrd A and Hrd B in cultivated (Hordeum vulgare L.) and wild (H. spontaneum C. Koch) barley from Iran (as a part of the Fertile Crescent)

Pomortsev A.A., Lyalina E.V.

Abstract

Polymorphism of hordeins encoded by the Hrd A and Hrd B loci was studied using starch gel electrophoresis in 63 landraces of cultivated barley (H. vulgare) and 43 accessions of wild barley (H. spontaneum) from Iran. We identified 35 and 32 Hrd A alleles of H. vulgare and H. spontaneum. The number of Hrd B alleles in these species was 55 and 48, respectively. The frequencies of alleles in the Hrd A and Hrd B loci in cultivated barley varied within the range of 0.0016–0.2816 (Hrd A) and 0.0016–0.1221 (Hrd B). The allelic frequencies in wild barley ranged within 0.0077–0.2170 and 0.0077–0.0815, respectively. Iranian accessions of H. spontaneum and H. vulgare shared some alleles; the number of common alleles was seven for the Hrd A locus and five for the Hrd B locus. These alleles were detected mainly in accessions of wild barley heterozygous or heterogenic for hordein-coding loci. We concluded that the regions close to Zagros mountain range cannot be considered as a barley domestication center. At the same time, Iranian H. spontaneum could have donated some hordein-coding loci to H. vulgare as a result of introgressions occurring because of spontaneous hybridization over the course of the spread of cultivated barley.

Russian Journal of Genetics. 2016;52(10):1034-1045
pages 1034-1045 views

The recovering of breeding achievements of Populus × leningradensis bogd. and Populus × newensis bogd. Based on microsatellite analysis

Lebedeva M.V., Levkoev E.A., Volkov V.A., Fetisova A.A., Navalikhin S.V., Shabunin D.A., Danilov Y.I., Zhigunov A.V., Potokin E.K.

Abstract

The genotyping of 75 trees from poplar plantations in St. Petersburg and Leningrad oblast was conducted with microsatellite markers to identify the elite clonal varieties developed by P.L. Bogdanov in the period of 1938–1965. The information about the varieties was lost. The authentic herbarium specimens of poplar clonal varieties preserved at the St. Petersburg State Forest Technical University were used as reference genotypes. According to the results of DNA fingerprinting, we identified the clonal plantations of Populus × newesis Bogd. and Populus × leningradensis Bogd. from the Kartashevskii forest district and the arboretum of the St. Petersburg State Forest Technical University. The identified elite poplar hybrids have a higher frost resistant and a higher growth rate. They are recommended for plantation cultivation in the northwest of Russia.

Russian Journal of Genetics. 2016;52(10):1046-1055
pages 1046-1055 views

Animal Genetics

New single nucleotide polymorphisms of androgen receptor gene (AR) in the Russian breed of Dzhalginsky Merino sheep

Trukhachev V.I., Krivoruchko A.Y., Skripkin V.S., Kvochko A.N., Kulichenko A.N., Kovalev D.A., Pisarenko S.V., Volynkina A.S., Selionova M.I., Aybazov M.M., Yatsyk O.A.

Abstract

This paper investigates the structure of androgen receptor gene (AR) in the Russian breed of Dzhalginsky Merino sheep. Polymorphisms of the gene were detected using NimbleGen sequencing technology (Roche, United States). Eight single nucleotide polymorphisms (SNPs) and two insertions were detected. Five of these SNPs (c.335T>G, c.339G>A, c.342T>C, c.2491-327T>A, and c.2491-325A>T) and both insertions were identified for the first time. Three SNPs and the insertions are located in the coding part of exon. Insertion c.336_337 is found in most of the animals of this breed and can be used as a genomic marker of the breed. Animals with mutant variant of SNP c.1496+15T>C have significantly lower live weight and body size compared with the wild type genotype. This SNP can be used as a genetic marker of meat production in marker-assisted selection.

Russian Journal of Genetics. 2016;52(10):1056-1061
pages 1056-1061 views

Seven genes of mitochondrial genome enabling differentiation of honeybee subspecies Apis mellifera

Ilyasov R.A., Poskryakov A.V., Nikolenko A.G.

Abstract

On the basis of comparative sequence analysis of 12 honeybee mitochondrial genes, seven genes enabling us to differentiate honeybees subspecies of the A, M, C, O evolutionary lineages were found. Applying comparative sequence analysis of ND2 gene mtDNA as an example on a statistically valid sample size, we showed a high level of differentiating ability of this gene and assumed that each of these seven genes probably can be used for differentiation of the subspecies within four evolutionary lineages.

Russian Journal of Genetics. 2016;52(10):1062-1070
pages 1062-1070 views

Differential expression of genes BPI, TAP1, SLA-1 and SLA-3 in Escherichia coli F18-resistant and sensitive Meishan post-weaning piglets

Yin X.M., Gan L.N., Qin W.Y., Sun S.Y., Zhu G.Q., Wu S.L., Bao W.B.

Abstract

The genetic basis and mechanism of sensitivity or resistance to the significant pathogen Escherichia coli F18 is not clear in native Chinese pig breeds and may differ from that in non-native breeds. Our previous research showed that the genes BPI, TAP1, SLA-1 and SLA-3 may play important roles in resistance to E. coli F18 in post-weaning piglets. This study was based on success in selecting and identifying full sib Chinese native Meishan breed post-weaning E. coli F18-resistant and sensitive piglets. Real-time PCR was used to detect expression levels of the genes BPI, TAP1, SLA-1 and SLA-3 in liver, spleen, thymus, lymph node, duodenum, and jejunum tissues between E. coli F18-resistant and -sensitive Meishan piglets. Only the BPI gene was obviously tissue specific; it was highly expressed in the duodenum and jejunum, but expressed at a low level in other tissues. The other three genes were expressed in all the studied tissues, and particularly highly in immune organs and intestinal tissues. The expression level of BPI in the duodenum of the resistant group was significantly higher than that in the sensitive group (P < 0.01). The SLA-3 expression level in the thymus of the resistant group was significantly higher than that in the sensitive group (P < 0.05). TAP1 and SLA-1 gene expression levels were generally higher in the resistant group than the sensitive group, but there were no significant differences. Genes BPI and SLA-3 play an important role in the processes of resistance to E. coli F18 in Meishan weaned piglets. We speculate that BPI protein may have a direct killing effect on Gram-negative bacteria such as E. coli strain F18 in the intestine; the resistance of Meishan weaned piglets to E. coli F18 is likely to be related to the upregulation of intestinal BPI. The upregulation of SLA-3 may increase the resistance of weaned piglets to E. coli F18 by regulation of the immune response.

Russian Journal of Genetics. 2016;52(10):1071-1076
pages 1071-1076 views

Human Genetics

Alu insertion-deletion polymorphism of COL13A1 and LAMA2 genes: The analysis of association with longevity

Erdman V.V., Nasibullin T.R., Tuktarova I.A., Somova R.S., Timasheva Y.R., Mustafina O.E., Karimov D.D.

Abstract

The distribution of allele and genotype frequencies of Alu(I/D) polymorphic sites in the COL13A1 and LAMA2 genes coding extracellular matrix protein subunits was characterized in an ethnically homogeneous group (Tatars from the Republic of Bashkortostan, Russia). It was established that the frequency of individuals with the COL13A1*D/*D genotype was higher in the senile age period. The LAMA2*I/*D genotype was predisposing to longevity among women. According to the observed results, the frequency of the LAMA2*I/*D genotype was increased in senile individuals older than 90 years. The observed associations can be explained on the basis of the contemporary view by the importance of Alu elements in gene expression regulation at transcriptional and post-transcriptional levels, the involvement of collagen and laminin in maintaining the structure and function of the extracellular matrix, and the relationship between the extracellular matrix state, pathological changes and aging.

Russian Journal of Genetics. 2016;52(10):1077-1085
pages 1077-1085 views

Population frequency and age of mutation G5741→A in gene NBAS which is a cause of SOPH syndrome in Sakha (Yakutia) Republic

Maksimova N.R., Nogovicina A.N., Kurtanov K.A., Alekseeva E.I.

Abstract

SOPH syndrome (Short stature with Optic nerve atrophy and Pelger–Huët anomaly syndrome, OMIM#614800) is an autosomal recessive hereditary disease characterized by the following main clinical symptoms: postnatal hypoplasia, proportionately short stature, facial dysmorphism, micromelia of feet and hands, limp and loose skin, optic nerve atrophy, and Pelger–Huët anomaly of neutrophils. For the first time, this disease was described in Yakuts. The molecular-genetic study showed that its cause in Yakuts is mutation G5741→A in gene NBAS. On the basis of disequilibrium analysis for linkage of ten microsatellite markers flanking the NBAS gene with the disease, the haplotype of the founder chromosome was determined. The age of the mutation in Yakutia was estimated to be about 804 ± 140 years. The frequency of heterozygous carriers of mutation G5741→A (R1914H) in gene NBAS was found, which averaged 13 per 1000 healthy Yakuts.

Russian Journal of Genetics. 2016;52(10):1086-1093
pages 1086-1093 views

Mathematical Models and Methods

Functional linear models for region-based association analysis

Svishcheva G.R., Belonogova N.M., Axenovich T.I.

Abstract

Regional association analysis is one of the most powerful tools for gene mapping because instead analysis of individual variants it simultaneously considers all variants in the region. Recent development of the models for regional association analysis involves functional data analysis approach. In the framework of this approach, genotypes of variants within region as well as their effects are described by continuous functions. Such approach allows us to use information about both linkage and linkage disequilibrium and reduce the influence of noise and/or observation errors. Here we define a functional linear mixed model to test association on independent and structured samples. We demonstrate how to test fixed and random effects of a set of genetic variants in the region on quantitative trait. Estimation of statistical properties of new methods shows that type I errors are in accordance with declared values and power is high especially for models with fixed effects of genotypes. We suppose that new functional regression linear models facilitate identification of rare genetic variants controlling complex human and animal traits. New methods are implemented in computer software FREGAT which is available for free download at http://mga.bionet.nsc.ru/soft/FREGAT/.

Russian Journal of Genetics. 2016;52(10):1094-1100
pages 1094-1100 views

Short Communications

Escherichia coli ydiO and ydiQRST genes encode components of acyl-CoA dehydrogenase complex of anaerobic fatty acid β-oxidation pathway

Gulevich A.Y., Skorokhodova A.Y., Debabov V.G.

Abstract

Escherichia coli open reading frames ydiO and ydiQRST were identified as genes encoding components of the acyl-CoA dehydrogenase complex of anaerobic fatty acid β-oxidation. Individual or concomitant inactivation of fadE gene, encoding known aerobic acyl-CoA dehydrogenase, and ydiO and/or ydiQRST genes did not affect cellular growth on glucose as a sole carbon source. Aerobic growth on sodium oleate was observed only for the cells with intact fadE gene. With an alternative electron acceptor, the cells possessing intact fadE gene demonstrated anaerobic growth on sodium oleate irrespective of the presence or absence of ydiO and ydiQRST genes. For the fadE-deficient mutants, anaerobic growth on sodium oleate was observed only for cells with intact ydiO and ydiQRST genes, while the fadE/ydiO and fadE/ydiQRST mutants failed to grow under the similar conditions.

Russian Journal of Genetics. 2016;52(10):1101-1104
pages 1101-1104 views

The search for new candidate genes involved in ovarian cancer pathogenesis by exome sequencing

Prokofyeva D.S., Mingajeva E.T., Bogdanova N.V., Faiskhanova R.R., Sakaeva D.D., Dörk T., Khusnutdinova E.K.

Abstract

Ovarian cancer is one of the most insidious of tumors among gynecological cancers in the world. BRCA1 and BRCA2 mutations are associated with high risk of ovarian cancer; however, they are causative only in a fraction of cases. The search for new genes would expand our understanding of the mechanisms underlying malignant ovarian tumors and could help to develop new methods of early diagnosis and treatment of the disease. The present study involved exome sequencing of eight DNA samples extracted from the blood of ovarian cancer patients. As a result of the study, 53057 modifications in one sample were identified on average. Of them, 222 nucleotide sequence modifications in DNA located in exons and splice sites of 203 genes were selected. On the basis of the function of these genes in the cell and their involvement in carcinogenesis, 40 novel candidate genes were selected. These genes are involved in cell cycle control, DNA repair, apoptosis, regulation of cell invasion, proliferation and growth, transcription, and also immune response and might be involved in development of ovarian cancer.

Russian Journal of Genetics. 2016;52(10):1105-1109
pages 1105-1109 views

Chromosomal localization of aromatic alcohol dehydrogenase fast-migrating isoenzyme Aadh1F (CAD1-F) gene in Triticum aestivum L. bread wheat

Konovalov A.A., Shundrina I.K., Karpova E.V., Goncharov N.P., Kondratenko E.Y.

Abstract

Differences in isoenzyme pattern of aromatic alcohol dehydrogenase, NADP-AADH or CAD, were found in the Triticum aestivum L. winter bread wheat cultivars by the method of electrophoresis in the starch gel. A standard three-component spectrum is present in the cv. Zitnica (former Yugoslavia); additional fact-migrating isoenzymes appear in the cv. Novosibirskaya 9 (Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, Russia). The presence of fast-migrating CAD isoenzymes is designated as FF phenotype; their absence, as 00 phenotype. Hybridological analysis was carried out; the excess of “null” genotypes was found in F2 progenies. Hybridization with nulli-tetrasomic lines of the chromosomes of the fifth homeologous group was conducted for the gene localization. The segregation analysis demonstrated the most probable localization of the CAD1-F gene in the chromosome 5A. The plants with FF and 00 genotypes differed in a number of chemical and anatomical traits, as well as in grain productivity. The results obtained are discussed in connection with the function of this enzyme in the wheat plant tissues.

Russian Journal of Genetics. 2016;52(10):1110-1116
pages 1110-1116 views

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