Russian Federation
Abstract. The role of allelic variability of inducible nitric oxide synthase (iNOS) is significant in the study of the resistance and susceptibility of animals to leukemia infection. After analyzing the literature data, it can be stated that in the iNOS gene, allele A (with genotype AA) is responsible for resistance to the leukemia virus, and allele B (with genotype BB) is responsible for susceptibility. This is due to the frequency of occurrence of alleles and their genotypes of the polymorphic marker AN13-1 of the inducibeal nitric oxide synthase gene. The iNOS gene is capable of producing a large amount of nitric oxide, compared to other isoforms. In turn, nitric oxide causes death or can stop the growth of pathogenic microorganisms, including viruses. The purpose of this work is to further study nitric oxide as an indicator for determining the resistance and susceptibility of animals to leukemia, as well as the selection of specific primers for PCR-PDRF used in genotyping. Methods. The iNOS gene sequence was analyzed and a pair of specific primers were selected and synthesized using the Vector NTI program. Scientific novelty of this work lies in the fact that we have selected specific primers that are important for the analysis of cattle genotyping by allelic variants of the polymorphic marker AH13-1 of the iNOS gene. Results. Based on this work, a pair of primers iNOSF_new and iNOSR_new, with a calculated annealing temperature of 52 °C, were selected and synthesized, giving an amplicon with a length of 186 bp. The amplicon contains a polymorphic site that distinguishes the A and B alleles. During PCR-RFLP, the following genotype-specific fragments are formed: AA-47/139 bp; AB -186/139/47 bp; BB-186 bp.
leukemia, bovine, interleukin, nitric oxide, oligonucleotides, resistance, susceptibility, polymorphic
1. Bartlett P. C., Ruggiero V. J., Hutchinson H. C., Droscha C. J., Norby B., Sporer K. R., Taxis T. M. Current developments in the epidemiology and control of enzootic bovine leukosis as caused by bovine leukemia virus // Pathogens. 2020. Vol. 9 (12). Article number 1058. DOI:10.3390/pathogens9121058.
2. Yang Y., Fan W., Mao Y., Yang Z., Lu G., Zhang R., Zhang H., Szeto C., Wang C. Bovine leukemia virus infection in cattle of China: Association with reduced milk production and increased somatic cell score // Journal of Dairy Science. 2016. Vol. 99 (5). P. 3688–3697. DOI: 10.3168/jds.2015-10580.
3. Petropavlovskiy M. V., Donnik I. M., Bezborodova N. A., Krivonogova A. S. Detection and immunobiological charcterizons of bovine leukeima virus in Russian Federation territory in dependence on geographical variations // Journal of Integrated OMICS. 2019. Vol. 9 (1). Article number 255. DOI: 10.5584/jiomics.v9i1.255.
4. Turkeev M. K., Mamanova M. K., Daugalieva A. T., Turgenbaev K. A., Kalisynov B. S. Epizootologiya i mery bor’by s leykozom krupnogo rogatogo skota v Respublike Kazakhstan [Epizootology and measures to combat bovine leukemia in the Republic of Kazakhstan] // Sbornik nauchnykh trudov KazNIVI. 2019. T. 65. Pp. 150–157. (In Russian.)
5. Kuzhebaueva U. Zh., Kakishev M. G., Koshemetov Zh. K. Epizooticheskaya situatsiya po leykozu krupnogo rogatogo skota v Zapadno-Kazakhstanskoy oblasti [Epizootic situation of bovine leukemia in the West Kazakhstan region] // Sovremennye vyzovy dlya biotekhnologii, veterinarii i meditsiny: Materialy mezhdunarodnoy nauchno-prakticheskoy konferentsii. Gvardeyskiy, 2020. Pp. 79–84. (In Russian.)
6. Gulyukin M. I., Stepanova T. V., Ivanova L. A., Kozyreva N. G., Shabeykin A. A., Kolomytsev S. A., Lopunov S. V., Barsukov Yu. I. Rasprostranenie i mery bor’by s leykozom krupnogo rogatogo skota v tsentral’nom federal’nom okruge [Distribution and measures to combat bovine leukemia in the Central Federal District] // Veterinariya i kormlenie. 2019. No. 6. Pp. 8–14. (In Russian.)
7. Hamada R., Metwally S., Polat M., Borjigin L., Ali A. O., Abdel-Hady A., Mohamed A., Wada S., Aida Y. A detection and molecular characterization of bovine leukemia virus in Egyptian dairy cattle // Frontiers in Veterinary Science. 2020. Vol. 7 (608). Pp. 1–13. DOI: 10.3389/fvets.2020.00608.
8. Aida Y., Murakami H., Takahashi M., Takeshima S. N. Mechanisms of pathogenesis induced by bovine leukemia virus as a model for human T-cell leukemia virus // Frontiers in Microbiology. 2013. Vol. 4 (328). Rp. 1–8. DOI: 10.3389/fmicb.2013.00328.
9. Lo C.-W., Borjigin L., Saito S., Fukunaga K., Saitou E., Okazaki K., et al. BoLA-DRB3 Polymorphism is associated with differential susceptibility to bovine leukemia virus-induced lymphoma and proviral load // Viruses. 2020. Vol. 12 (3). Article number 352. DOI: 10.3390/v12030352.
10. Zyrianova I. M., Koval’chuk S. N. Bovine leukemia virus pre-miRNA genes’ polymorphism // RNA BIOLOGY. 2018. Vol. 15 (12). Rp. 1440–1447. DOI: 10.1080/15476286.2018.1555406.
11. Takeshima S., Ohno A., Aida Y. Bovine leukemia virus proviral load is more strongly associated with bovine major histocompatibility complex class II DRB3 polymorphism than with DQA1 polymorphism in Holstein cow in Japan // Retrovirology. 2019. Vol. 16. Article number 14. DOI: 10.1186/s12977-019-0476-z.
12. Gil’manov Kh. Kh., Vafin R. R., Karimova R. G., Tyul’kin S. V. Sposob provedeniya PTsR-PDRF dlya genotipirovaniya krupnogo rogatogo skota po allel’nym variantam polimorfnogo markera AH13-1 gena iNOS [The method of PCR-PDRF for cattle genotyping by allelic variants of the polymorphic marker AH13-1 of the alien gene] // Veterinary, Zootechnics and Biotechnology. 2018. No. 4. Pp. 22–28. (In Russian.)
13. Mishchenko V. A., Petrova O. N., Karaulov A. K., Mishchenko A. V. Problema leykoza krupnogo rogatogo skota [The problem of bovine leukemia]. Vladimir: FGBU “VNIIZZh”, 2018. 38 p. (In Russian.)
14. Aytnazarov R. B., Ignat’eva E. V., Agarkova T. A., Dvoeglazov N. G., Osipova N. A., Khramtsov V. V., Yudin N. S. Assotsiatsiya odnonukleotidnogo polimorfizma rs110861313 v mezhgennom rayone khromosomy 23 s razvitiem leykoza u krupnogo rogatogo skota cherno-pestroy porody [Association of single nucleotide polymorphism pc110861313 in the intergenic region of chromosome 23 with the development of leukemia in black-and-white cattle] // Vavilov Journal of Genetics and Breeding. 2019. No. 23 (8). Pp. 999–1005. DOI: 10.18699/VJ19.576. (In Russian.)
15. Chichinina S. V. Rol' allel'noy variabel'nosti genov tsitokinov v formirovanii rezistentnosti krupnogo rogatogo skota k leykozu: dis. … kand. biol. nauk: 16.00.03, 03.00.23 [The role of allelic variability of cytokine genes in the formation of resistance of cattle to leukemia: dissertation ... candidate of biological sciences: 16.00.03, 03.00.23]. Novosibirsk, 2005. 107 p. (In Russian.)
16. Yudin N. S., Podkolodnyy N. L., Agarkova T. A., Ignat’eva E. V. Prioritizatsiya genov, assotsiirovannykh s patogenezom leykoza u krupnogo rogatogo skota [Prioritization of genes associated with the pathogenesis of leukemia in cattle] // Vavilov Journal of Genetics and Breeding. 2018. No. 22 (8). Pp. 1063–1069. DOI: 10.18699/VJ18.451. (In Russian.)
17. Gil’manov Kh. Kh. Genotipirovanie krupnogo rogatogo skota po genam, opredelyayushchim ustoychivost’ k leykozu, i genoidentifikatsiya ego etiologicheskogo agenta: dis... kand.biol.nauk [Genotyping of cattle by genes determining resistance to leukemia and genoidentification of its etiological agent: dissertation ... candidate of biological sciences]. Kazan, 2019. 163 p. (In Russian.)
18. Ivanov D. V. Immunologiya. Immunodefitsity zhivotnykh: uchebnoe posobie [Immunology. Animal immunodeficiency: a textbook]. Bryansk: Bryanskiy GAU, 2019. 154 p. (In Russian.)
19. Stone M. J., Hayward J. A., Huang C., Huma Z. E., Sanchez J. Mechanisms of Regulation of the Chemokine-Receptor Network // International Journal of Molecular Sciences. 2017. Vol. 18 (2). Article number 342. DOI: 10.3390/ijms18020342.
20. Arsent’eva N. A., Semenov A. V., Zhebrun D. A., Vasil’eva E.V., Totolyan A.A. Rol’ khemokinovogo retseptora CXCR3 i ego ligandov pri nekotorykh immunopatologicheskikh sostoyaniyakh [The role of the chemokine receptor CXCR3 and its ligands in some immunopathological conditions] // Medical Immunology. 2019. T. 21. No. 4. Pp. 617–632. (In Russian.)
21. Cheng Y., Huang C., Tsai H. Relationship of bovine NOS2 gene polymorphisms to the risk of bovine tuberculosis in Holstein cattle // Journal of Veterinary Medical Science. 2016. Vol. 78 (2). Pp. 281–286. DOI: 10.1292/jvms.15-0295.
22. Widdison S., Ashley G. R., Howard C. J., Coffey T. J. Characterisation of bovine inducible nitric oxide synthase // Veterinary Immunology and Immunopathology. 2007. Vol. 117 (3-4). Pp. 302–309. DOI: 10.1016/j.vetimm.2007.01.016.