1 Подільський державний аграрно-технічний університет, вул. Шевченка, 13, м. Кам'янець-Подільський, 32316, Україна 2 Інститут розведення і генетики тварин імені М. В. Зубця НААН, вул. Погребняка, 1, с. Чубинське, Бориспільський р-н, Київська обл., 08321, Україна LL (0,964) і, відповідно, алеля L (0,982), та відсутність генотипу VV. області (стадо № 2). Спектр алелів генів гормону росту (GH), бета-лактоглобуліну (βLG), капа-казеїну (CSN3), тиреоглобуліну (TG5) і калпаїну (CAPN) вивчали за допомогою ПЛР-ПДРФ. Дослідження генетичного поліморфізму генів двох популяцій української сірої породи показує, що за геном капа-казеїну виявлено переважання генотипу АВ (0,476) при прямо протилежному розпо-ділі частот алелів А і В. За геном тиреоглобуліну простежується подібний розподіл алелів С і Т. Ви-явлено переважання генотипу СТ (0,571) у тварин стада № 1 і надлишок гомозигот для корів стада

Buffalo breeding in Ukraine is an ancient traditional branch of animal husbandry of the Crimean Tatars and Rusyns of Transcarpathia. Basically, "Ukrainian" buffaloes belong to the river buffalo (Bubalus bubalis) and are bred for dairy and meat production. Polymorphism of genes associated with dairy productivity will allow breeding buffaloes taking into account the “desired” genotypes in relation to economically useful traits. The paper dwells on studying allelic polymorphisms of beta-casein (β-CN), kappa-casein (CSN3) and beta-lactoglobulin (β-LG) genes in population of water buffaloes bred in Ukraine using PCR followed by restriction hydrolysis of the formed fragments (PCR-RFLP). Results of study of the “Ukrainian” population of water buffaloes are discussed, namely: the frequency of genotypes and alleles at the loci of beta-casein, kappa-casein and beta-lactoglobulin genes. Amplified fragment β-CN with the length of 121 bp was digested with DdeI restriction enzyme. A feature of the allelic spectrum of the beta-casein gene (β-CN) in the studied population was absence of A1 allele. All animals carried the β-CNA2A2 genotype of beta-casein gene, respectively, β-CNA2 allele frequency was 1.0. For the CSN3 gene, an amplified fragment in 273 bp was digested with HinfI restriction enzyme. A 100% predominance of animals with the most preferred homozygous CSN3BB genotype was revealed. During the β-LG gene study process, an amplified fragment with a size of 247 bp was digested with HaeIII. It has been determined that the most frequent was allele βLGA and genotype βLGAA of beta-lactoglobulin gene (0.96 and 0.92, respectively). Heterozygous β-LGAB genotype is present in 8 % of buffaloes. The research results are of interest in the field of molecular genetic analysis of the buffalo genome, which are the source of specific properties. The data obtained can be useful for preserving and increasing the genetic diversity of the “Ukrainian” population of water buffaloes, as well as for obtaining valuable products from buffaloes.

At the current stage of genetic studies of cattle, more and more attention is being drawn to autochthonous breeds. Native cattle have a number of prominent phenotypic traits and have preserved unique genes and their combinations lost by modern commercial breeds, which would be valuable to use in selective programs. We surveyed polymorphism of the Ukrainian autochthonous Gray breed according to alleles of exon 2 of the BoLA-DRB3 gene. The uniqueness of the gene lies in the broad variability of its allele variants. Significant informativeness at DNA level is quite important for genetic studies. We surveyed allele polymorphism using the PCR-RLFP method on DNA isolated from 88 samples of blood of cows and 5 samples of sperm. We identified 28 alleles, of which 23 variants were nomenclature ones and 5 (jba, *jab, *jbb, *nad and *nda) were “without established nomenclature”, their share accounting for 8.9%. Four alleles *06, *12, *16 and *jba had a frequency above 5% and occupied 69.9% of the breed’s allele fund overall. The commonest allele was BoLA-DRB3.2*16 (44.1%). In total, we found 40 genotypes. Considering the significant dominance of variant *16, as expected, 5 genotypes with its inclusion occurred: *16/*16, *12/*16, *06/*16, *16/*24 and *jba/*16. It was present in the genotype of two out three studied animals. Parameters of heterozygosity, effective number of alleles, Shannon and Pielou indices indicate that Ukrainian Gray cattle are characterized by lowest level of genetic variability and biodiversity according to the BoLA-DRB3 gene compared with other breeds. Due to significant dominance of allele *16, the breed has no inbred motifs. We noted deviation toward increase in homozygosity without deviations from the norm of the distribution according to Hardy-Weinberg equilibrium. The obtained results will be used for genetic-populational programs with the purpose of improving the genetic potential of cattle breeds in terms of economically beneficial traits and diseases of cattle.

Today conservation of biodiversity is one of the most important priorities in the world. Polymorphism of farm animals is a key to successful breeding; it provides animal adaptation to environmental changes. Impoverishment of genetic resources of farm animals can lead to various negative consequences, such as significant decrease of the effectiveness of selection, existing breeds will not be able to successfully resist to infectious agents evolving constantly; valuable material for the study and analysis of breed origin will be lost. Local breeds created by national selection are valuable genetic resources. Lacking high performance in most cases, they are usually characterized by high resistance to various diseases. Ukrainian Grey cattle, as a representative of the local native breeds, are an interesting object of population research in respect not only of adaptive characteristics, but also genetic mechanisms providing phenotypic expression of certain features of productivity. Recent advances in molecular genetics have made it possible to identify genes associated with qualitative and quantitative characteristics of cattle. The most informative one in this regard is the DNA marker systems based on structural analysis of polymorphisms of genes involved in formation and operation of economically useful traits. The most common potential DNA markers of characteristics of cattle productivity include genes of growth hormone (bGH), beta-lactoglobulin (βLG), thyroglobulin (TG5), calpain (CAPN). BGH gene is an important regulator of somatic growth of animals and has lactating and fat-mobilizing effects. CSN3 gene is linked to milk protein ability and technological properties of milk. Different allelic variants of βLG gene are associated with high content casein and whey proteins in milk, fat percentage and positive impact on milk production. Milk products and falsification detection of milk are controlled by this gene. Its role has been proved in antimicrobial activity to pathogens of mastitis. TG5 gene is a precursor of tyreoid hormones such as triiodothyronine and tetraiodothyronine participating in formation of fat cells and the formation of meat marbling. CAPN gene is involved in proteolysis during ripening of meat and leads to higher tenderness of meat. The aim was to investigate the allelic polymorphism of genes of growth hormone, beta-lactoglobulin, thyroglobulin and calpain in Ukrainian Grey cattle. Blood samples (n = 136) from Ukrainian Grey cattle at "Markeyevo" breeding farm (Kherson region) were studied. Molecular genetic studies were conducted at Laboratory of Genetics of Institute of Animal Breeding and Genetics nd. a. M.V. Zubets of NAAS. DNA isolation from whole blood was performed using standard commercial kit "DNA-Sorb-B" (produced by AmpliSens, Central Research Institute of Epidemiology, Russia). Polymorphisms of genes of GH, βLG5, TG and CAPN1530 was investigated by PCR-RFLP. Electrophoretic separation of restriction fragments of DNA was performed in 1.5% and 2% agarose gels in tris-borate electrophoresis buffer. The results of DNA testing of beta-lactoglobulin locus for A- and B-allele variants in animals of Ukrainian Grey breed found that most of the cows were carriers of homozygous BB genotype of βLG gene. It was determined at every second investigated animals. Homozygous AA genotype was found only at two cows or it is 4%. AB genotype was represented at 45% of animals. Thus, the study found a significant prevalence of frequencies B allele over A allele (0.736 and 0.264, respectively), which leads to overwhelming homozygotisation of one of these alleles. B allele in beta-lactoglobulin gene is associated with high content of casein protein in milk and a high percentage of fat. The values of expected and observed heterozygosity at this gene were not significantly different. The study of gene polymorphism of thyroglobulin found that at the population of the represented breed, the most often seen heterozygous genotype was CT, carriers of which were 57% of the animals. In homozygous state C allele was in 31% of the animals, and T allele – only in 11%. Regarding the level of heterozygosity, we received that actual heterozygosity exceeded theoretically expected one. However, difference is statistically insignificant. Consequently, for thyroglobulin gene Ukrainian Grey cattle belong to breeds that carry the highest frequency of desired C allele for marbling of meat. The analysis of gene polymorphism of bGH indicates the absence of the animals with genotype VV in the investigated sampling, low percentage of heterozygotes (3%) and a significant portion of homozygotes for L allele (98%). Significant differences on levels of actual and expected heterozygosity for somatotropin gene we haven’t found. One of the markers of quality characteristics of meat productivity of cattle is CAPN gene. In general, the animals of Ukrainian Grey breed are characterized by the absence of polymorphism for calpain gene. All the studied animals were carriers of homozygous genotype for preferred G allele (1,0). This feature of the genetic structure of the studied population by calpain gene, animals of which are reproduced in a small array, shows breed-specific high genetic potential by quality characteristic of meat productivity, namely, the tenderness of meat. It was revealed that the population of Ukrainian Grey breed was characterized by low level of polymorphism for genes which cause milk production and meat quality indicators. Thus, homozygous genotype BB dominated for beta-lactoglobulin gene; homozygous LL genotype (98%) was also significantly prevalent and VV genotype was not detected for growth hormone gene; only genotype GG was generally found for calpain gene. And heterozygous CT genotype (57%) dominated only for TG5 locus of thyroglobulin gene.

The history of the origin and domestication of farm animals has always interested mankind. However, these issues are covered in the literature in great detail only from the time when herds of domestic animals have already formed. Most often, the genesis of individual species, the original forms that formed the basis of domestication, remain unclear. [2] An example is the history of domestication of the horse, as the horse played a central role among other domestic animals in the development of human society. In the study of mammal fauna of the Pleistocene-Holocene of Europe there is a problem of studying the origin of the domestic horse Eguus cabalus L., ie, the establishment of wild ancestors of domesticated breeds, place, time and process of their domestication. Analysis of literature data on paleontological and archaeological finds in Ukraine showed that most researchers believe that the first domesticated horses began to recognize horses, the remains of which were found during archaeological excavations of the settlement of the third millennium BC. BC in Botai (Northern Kazakhstan), but from which taxon the opinions of scientists differ. Some believe that it could be Tarpan, however, there is an opinion that a large horse could not come from a small tarpan and Przewalski's horse. Therefore, preference was given to the hypothesis of the origin of the domestic horse from the ancient Pleistocene. At present, the problem of the origin of the domestic horse does not go beyond hypotheses and assumptions, and this is primarily due to the slight difference between the bones of the domestic and wild horse. The plasticity of the skeleton of the genus Eguus is very weak and this explains the problems faced by paleontologists in trying to develop the evolutionary history of horses. Thus, to understand the processes of domestication of this animal, in addition to archaeological and paleontological research methods, it is necessary to use tools from other fields of science, such as molecular genetic analysis of DNA samples. One of the variants of test systems for studying genetic polymorphism is the use of ISSR markers, which allow to analyze DNA fragments and make certain phylogenetic connections in the studied groups. In the laboratory of genetics of the Institute of Breeding and Genetics of Animals named after M.V.Zubets NAAS began research in the field of paleogenetics, namely – the study of the molecular genetic component in the fossils of ancient members of the genus Eguus using ISSR-markers. Inverted repeats are of particular interest because they are unevenly distributed throughout the genome and do not require prior knowledge of the nucleotide sequence of the test DNA. A significant point in the selection of research methods for us was that intermicrosatellite polymorphism is used to study interspecific and intraspecific genetic variability. It is believed that DNA fragments obtained by ISSR analysis can be species- and breed-specific, and this method is widely used by researchers in the study of breed groups. The purpose of our work is to develop a new method of DNA isolation from fossil remains (bones) of ancient horses and the production of ISSR-PCR with isolated DNA samples in the laboratory of genetics IRGT. M.V.Zubets NAAS according to the available reagents and existing protocols. The research was carried out on samples of fossil bones of horses of the Pleistocene period (about 10 thousand years BC). One bone was found in the village. Beeches of Zhytomyr region in a career. Excavations were carried out in 1960, the metacarpal bone (os. Tarsicentral). Another bone was found in Novgorod-Siversky, Chernihiv region. in a career. Excavations were conducted by Boriskovsky PI in 1935. A tooth found in the village of Tarpan was used to study a wild tarpan horse (4.5 thousand years BC). Skibnytsia, Trostyanets district, Vinnytsia region. Excavations were conducted in 1959 by VM Danylenko. The paleontological material for the study was provided by the Kyiv National Museum of Natural History of the National Academy of Sciences of Ukraine, Department of Paleontology. As a result of this work for the first time in the Department of Genetics and Biotechnology IRGT. Research on paleogenetics has been started by M.V.Zubets. We optimized the method of extracting genetic material from fossils and obtained DNA from the bones of a horse of the Pleistocene period (about 10 thousand years BC) and the tooth of a wild horse tarpan (4.5 thousand years BC). Also, the optimal conditions for PCR were selected to work with DNA obtained from fossil remains, to study polymorphism with ISSR markers, and electrophoregrams of amplification products were obtained.