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Aims: Caseins are among the main milk proteins that determine many of its properties. Bovine kappa-casein (CSN3) is associated with the qualitative composition of milk, as well as with the quality of cheese obtained from this milk. The rs43703016 single-nucleotide substitution (g.88532332A>C; Asp148Ala) in exon 4 of the bovine CSN3 gene plays an important role in the production of quality hard cheeses. Various methods for the DNA testing of this substitution have been developed in the last three decades. Emergent DNA technologies provide an opportunity to modernize methods of genotyping single-nucleotide polymorphisms. Results: We have developed and verified a method to differentiate A/C alleles of the rs43703016 substitution in the bovine CSN3 gene by real-time PCR using allele-specific fluorescent probes. Conclusion: Our new method allows fast genotyping of animals, and may be used for selection of cows carrying the CC genotype, which determines good cheese-making properties of milk.

Allele frequencies of three substitutions in the CCK (rs42891945 and rs42891946) and CCKBR (rs42670352) genes were identified in three Russian cattle breeds: Holstein (dairy), Bestuzhev (dual-purpose), and Kalmyk (beef). The most promising for further associative studies is the rs42670352 substitution in the CCKBR gene.

The 32-bp deletion ( CCR5del32 mutation) in the CCR5 (chemokine (C-C motif) receptor 5) gene, encoding CCR5 chemokine receptor, is one of the factors determining natural resistance to human immunode- ficiency virus (HIV-1) infection. In the present study, the samples of Russians ( n = 102), Tuvinians ( n = 50), and HIV-infected individuals ( n = 107) were examined for the presence of CCR5del32 mutation in the CCR5 gene. The CCR5del32 allele frequency in Russians and Tuvinians constituted 7.84 and 2%, respectively. Among HIV-1 infected individuals, two groups, of macrophage-tropic HIV-1 strain- and T-cell-tropic HIV-1 strain- infected were distinguished. The CCR5del32 allele frequency in the first group (6.45%) was lower than in the second one (8.73%). Statistical treatment of the HIV-1 infected individuals typing data showed that the differ- ence in the CCR5del32 allele frequencies between the groups of sexually (macrophage-tropic) and parenterally (T-cell-tropic) infected individuals observed was within the limit of random deviation.

Imprinted genes play important roles in the mammalian development. In the parthenogenetic embryos (PE), there is only expression of maternally expressed genes. Therefore, PEs are appropriate experimental models to study genomic imprinting controlling mechanisms. The maternally expressed H19 and paternally expressed Igf2 are reciprocally imprinted genes in normal embryos. Here, we studied effect of transforming growth factor alpha (TGFα) treatment in vitro (10 ng/ml at the morula stage) on the expression of Igf2/H19 locus in mice PE (9.5 days of gestation, 25 somites) and their placentas (PP). Using RT-PCR, we showed that TGFα reactivated maternally imprinted Igf2 gene in parthenogenetic embryos and placentas. In spite of similar Tgfα expression in the preimplantation stages, its expression in the 9.5-day parthenogenetic embryos is significantly less than in normal embryos (NE). In our experiments, it was shown that reactivation of Igf2 gene occurred independently of H19 gene. In vitro TGFα treatment of mouse PE reactivated paternally expressed Igf2 gene in the PE and PP. In the PE and PP, both Igf2 and H19 were expressed. It seems that TGFα can play an important role as modulator of the Igf2/H19 locus.

The exon–intron structures of the human, rat and mouse ABLIM2 gene were determined in silico. The experimental verification resulted in the revealing of two mRNA isoforms of the ABLIM2 gene. The isoforms a and b contained 20 exons and 18 exons, respectively. The highest expression of both isoforms was observed in rat brain and eye and in mouse embryos. The 5V -UTR region of the ABLIM2 gene was 127 bp in rat and mouse, but in human, it was 65 bp. The site of polyadenylation was shown to be present at a distance of 682 bp from the stop-codon in human and rat and 684 bp in mouse. The in silico analysis of the gene 5V was performed. The high density of brain and CNS specific transcription factors’ binding sites in the promoter region was shown for all three organisms. The comparison of the amino acid sequences of the human ABLIM2 and ABLIM1 proteins showed that the number and arrangement of domains (four LIM-domains in the Nend region and the C-end VHP-domain) were similar. The structure of the ABLIM2 proteins was similar in all three organisms. On the basis of our data, it was assumed that the ABLIM2 protein was necessary for the normal functioning of neurons. D 2005 Elsevier B.V. All rights reserved.

The structure was described for human gene CKAP2, which codes for the cytoskeleton-associated protein 2, is at the boundary of chromosome regions 13q14.3 and 13q21.1, and is rearranged in various tumors. The CKAP2 exon–intron structure was established by collating the nucleotide sequences of the cDNA and genomic clone AL359513 (GenBank) and analyzing the gene sequence with the GENSCAN program. The results were verified by amplifying gene fragments. CKAP2 comprises nine exons ranging 70–1442 bp and is about 22 kb in size (regulatory regions included). The CKAP2 promoter contains CCAAT (–39...–33) rather than the canonical TATA box, and harbors nine binding sites for six transcription factors. Thus, CKAP2 possesses all structural elements characteristic of eukaryotic genes. The results may be used to study the CKAP2 expression in normal and tumor cells in order to elucidate its role in carcinogenesis.

The dopaminergic system plays a major role in migraine. Dopamine beta hydroxylase (DBH) is responsible for maintaining dopamine-to-norepinephrine ratio implicated in migraine pathophysiology. We aimed to look for association of polymorphisms in dopaminergic genes in genetic susceptibility to migraine in Russian population. In the present study DBH polymorphisms rs1611115 was selected. The aim of this study was to determine whether the polymorphisms rs1611115 in DBH gene influenced any particular symptoms of the disease.

Using a new pEnLox vector employed to generate gain-of-function mutants in Arabidopsis thaliana, the AtFAS4 mutant has been obtained and analyzed. The mutant is characterized by super-expression of the At1g33390 gene, which leads to the occurrence of a mutant phenotype – stem fasciation. The level of expression of the AtFAS4 gene in normally developing A.thaliana plants is extremely low thus accounting for almost complete absence of information on EST’s of this gene. The generated AtFAS4 mutant has permitted full-length cDNA of the At1g33390 gene to be obtained and analyzed for the first time.

By computer sequence analysis, the exon-intron structure of the human WASF4 gene was determined and the binding sites for transcription factors were detected in the promoter region. WASF4 was most homologous to human WASF2. WASF4 homologs were found in the macaque and chimpanzee genomes but not in other vertebrate genomes. Expression of WASF4 was not detected in human tissues.

Homozygous deletions or loss of heterozygosity (LOH) at human chromosome band 3p12 are consistent features of lung and other malignancies, suggesting the presence of a tumor suppressor gene(s) (TSG) at this location. Only one gene has been cloned thus far from the overlapping region deleted in lung and breast cancer cell lines U2020, NCI H2198, and HCC38. It is DUTTI (Deleted in U Twenty Twenty), also known as ROBO1, FLJ21882, and SAX3, according to HUGO. DUTTI, the human ortholog of the fly gene ROBO, has homology with NCAM proteins. Extensive analyses of DUTTI in lung cancer have not revealed any mutations, suggesting that another gene(s) at this location could be of importance in lung cancer initiation and progression. Here, we report the discovery of a new, small, homozygous deletion in the small cell lung cancer (SCLC) cell line GLC20, nested in the overlapping, critical region. The deletion was delineated using several polymorphic markers and three overlapping PI phage clones. Fiber-FISH experiments revealed the deletion was approximately 130 kb. Comparative genomic sequence analysis uncovered short sequence elements highly conserved among mammalian genomes and the chicken genome. The discovery of two EST clusters within the deleted region led to the isolation of two noncoding RNA (ncRNA) genes. These were subsequently found differentially expressed in various tumors when compared to their normal tissues. The ncRNA and other highly conserved sequence elements in the deleted region may represent miRNA targets of importance in cancer initiation or progression. Published 2006 Wiley-Liss, Inc.(dagger).

Results The genotype frequencies of -1021C/T polymorphism DBH gene did not differ between the MOH and other groups. In contrast, we found that the presence of the -1021T allele was significantly associated with the monthly drug consumption and LDQ total score in the MOH group. We also found an interaction between the presence of DBH -1021T genotype and efficiency serotonin-norepinephrine reuptake inhibitors in patient with MOH. Conclusions