Interferons have a long history of use in healthcare for the treatment of viral diseases. In veterinary practice, however, the introduction of interferons for the prevention and treatment of cattle is just beginning. Foreign interferon-based drugs are already available on the Kazakhstan market, and demand for them increases significantly during seasonal infections or outbreaks of viral diseases. In this regard, establishing the production of recombinant IFN is an urgent task for Kazakhstan. Any production of biological preparations begins with research. The proposed project represents the first stage in developing domestic production of recombinant interferons for the needs of animal husbandry. Within this project, four recombinant cattle IFNs will be obtained, their antiviral activity assessed, and their biological properties studied. Producer strains will be deposited, methods for producing recombinant IFNs preserved, and the stability of their activity during storage will also be evaluated. In the future, this will enable commercialization of the technology. The project has significant scientific novelty, as it involves comparing the biological properties of different interferons (various subtypes of IFN type I) under the same conditions. This will allow us to determine which IFN is most effective against specific viruses that cause cattle diseases.
To obtain recombinant type I interferons (different subtypes) from cattle and study their antiviral activity against cattle viruses.
The main idea of the project is to obtain recombinant bovine type I interferons (different subtypes) and compare their antiviral activity against viruses that cause bovine diseases. As a result of the project, at least four recombinant bovine type I interferons will be produced, with specific activities for inhibiting the replication of bovine viruses measured. This will allow for the selection of the most effective recombinant interferons for the development of promising antiviral veterinary drugs. Throughout the work, protocols for the production, purification, and refolding of the four interferons will be optimized. Subsequently, the most active substances and methods for their production will be utilized to commercialize the technology for producing bovine interferons in Kazakhstan.
Kairzhanova Alma Duysenbaykyzy, 35 years old, is a PhD and research associate at the Laboratory of Applied Genetics of the National Centre of Biotechnology. She has 13 years of scientific experience. Her research interests include molecular epidemiology, the development of diagnostic test systems, epizootology, microbiology, molecular biology, and next-generation sequencing methods. Her Hirsch index is 2 (Scopus). Her main tasks involve project management, writing reports, scientific articles, and experimental design.
https://www.webofscience.com/wos/author/record/2098064
https://orcid.org/0000-0002-9864-2700 –
https://www.scopus.com/authid/detail.uri?authorId=56737034300.
1. Kuibagarov M., Kairzhanova A., Vergnaud G., Amirgazin A., Lukhnova L., Izbanova U., Ramankulov Y., Shevtsov A. Draft Genome Sequence of the Strain Francisella tularensis subsp. mediasiatica 240, Isolated in Kazakhstan // Microbiol. Resour. Announc. 2020 Aug 27;9(35):e00766-20. IF-0.7, CiteScore -1.5, https://doi.org/10.1128/mra.00766-20 Web of Science Q4.
2. Shevtsov V., Kairzhanova A.,Shevtsov A., Shustov A., Kalendar R., Abdrakhmanov S., Lukhnova L., Izbanova U., Ramankulov Y., Vergnaud G. (2021) Genetic diversity of Francisella tularensis subsp. holarctica in Kazakhstan // PLoS Negl Trop Dis. 15(5):e0009419. CiteScore -6.4, IF- 3.4. doi: 10.1371/journal.pntd.0009419. Web of Science Q1.
3. Shevtsov A., Ramanculov E., Shevtsova E., Kairzhanova A., Tarlykov P., Filipenko M., Dymova M., Abisheva G., Jailbekova A., Kamalova D., Chsherbakov A., Tulegenov S., Akhmetova A., Sytnik I., Karibaev T., Mukanov K. Genetic diversity of Brucella abortus and Brucella melitensis in Kazakhstan using MLVA-16 // Infection, Genetics and Evolution 34 (2015): 173-180. https://doi.org/10.1016/j.meegid.2015.07.008 IF-2.6. Web of Science Q1
4. Shevtsov A., Izbanova U., Amirgazin A., Kairzhanova A., Dauletov A., Kiyan V., Vergnaud G. Genetic Homogeneity of Francisella tularensis subsp.mediasiatica strains in Kazakhstan//Pathogens. 2024 Jul 12;13(7):581. IF-2.6 https://doi.org/10.3390/pathogens13070581 Web of Science Q2.
Mukanov Kasym Kasenovich, Doctor of Veterinary Sciences, is a Professor and Chief Researcher at the Laboratory of Applied Genetics of the National Center for Biotechnology. He has 45 years of scientific experience. His research interests include virology, immunology, the study of the epidemiology of infectious diseases and the genetic diversity of pathogens, as well as the development of diagnostic systems, epizootology, and microbiology. As part of the project, he will participate in creating strains of microorganisms that produce recombinant interferons and developing methods for their purification. He is the author of 23 articles in foreign publications with an impact factor and holds 30 security documents for inventions. His Hirsch index is 7 (Scopus).
https://www.webofscience.com/wos/author/record/1371888
https://www.scopus.com/authid/detail.uri?authorId=56340590800
https://orcid.org/0000-0002-0502-9238.
Shevtsov Aleksandr Borisovich, PhD in Biology, is an Associate Professor and Head of the Laboratory of Applied Genetics at the National Centre for Biotechnology, with 22 years of scientific experience. His research field includes Sanger sequencing and next-generation sequencing (NGS) using Ion Torrent and MiSeq, MLVA typing, cloning, primer design, real-time PCR, and bioinformatics analysis. As part of the project, he will participate in bioinformatics analysis and primer selection for PCR amplification of the interferon gene, as well as writing reports, scientific articles, and experimental design. His Hirsch index is 10 (Scopus).
https://www.webofscience.com/wos/author/record/1632219
https://www.scopus.com/authid/detail.uri?authorId=57201604158
https://orcid.org/0000-0002-0307-1053.
Shustov Aleksandr Vyacheslavovich, Ph.D. in Biology, is the Head of the Genetic Engineering Laboratory at the National Centre of Biotechnology, with 32 years of scientific experience. His research area includes genetic engineering of microorganisms, virology, bioengineering of expression systems in mammalian cells and bacteria, and the production of recombinant proteins. As part of the project, he will participate in creating genetic constructs containing the bovine interferon gene, as well as optimizing the production and purification of recombinant proteins. His Hirsch index is 12 (Scopus).
https://www.webofscience.com/wos/author/record/430969
https://www.scopus.com/authid/detail.uri?authorId=57211989685
https://orcid.org/0000-0001-9880-9382.
Ryskeldina Anara Zhankozhaevna, 28 years old, PhD student, junior researcher of Applied Genetics Laboratory of NCB. Experience of scientific work for 6 years. Area of research: study of epidemiology of infectious diseases and genetic diversity of pathogens, Sanger sequencing, development of diagnostic test systems based on PCR and ELISA, obtaining recombinant proteins. Within the framework of the project he will take part in the development of a protocol for the production, purification and refolding of bovine interferons. Hirsch Index (Scopus) =2
https://www.webofscience.com/wos/author/record/56097356
https://www.scopus.com/authid/detail.uri?authorId=57218871763
https://orcid.org/0000-0002-7100-2711.
Tursunbay Naila Erlankyzy, 24 years old, holds a Master’s degree and works as a laboratory assistant at the Applied Genetics Laboratory of the National Central Hospital, with 2 years of scientific experience. Her research field includes the study of the epidemiology of infectious diseases and the development of PCR methods for diagnosing blood parasitic and infectious diseases in humans and animals. As part of the project, she will participate in the isolation and purification of target proteins, as well as obtaining substances and creating original preparative forms of recombinant interferon drugs.
https://www.webofscience.com/wos/author/record/64038358
https://www.scopus.com/authid/detail.uri?authorId=59304069500
https://orcid.org/0000-0002-6604-4247.
Dauletov Ayan Erbolatovich, 23 years old, Master’s degree, laboratory assistant at the Applied Genetics Laboratory of the National Central Hospital. 2 years of scientific experience. Area of research: study of infectious diseases of animals and their genetic diversity, Sanger sequencing, development of PCR test systems. As part of the project, he will participate in the production and purification of the target protein, ELISA and Western blot.
https://www.webofscience.com/wos/author/record/60852457
https://orcid.org/0000-0003-1906-6889.
Kasen Amirkhan Mauliuly, 23, laboratory assistant at the Applied Genetics Laboratory of the National Central Hospital. 2 years of scientific experience. Research area: genotyping, sequencing, studying the genetic diversity of various pathogens. As part of the project, he will take part in the colonisation of target fragments, purification and refolding of the recombinant protein.
https://www.webofscience.com/wos/author/record/50933821
https://orcid.org/0000-0002-8488-0863.
Collections of nucleotide sequences of bovine interferon type I genes deposited in international databases of genetic information and the in silico construction of sequences for expression systems in E.coli have been compiled. In the course of the work, an expanded collection of nucleotide sequences of type I interferon genes in cattle was formed, including representatives of all major subtypes of IFN-α, IFN-β and IFN-τ.
Primers and chemical synthesis of codon-optimized sequences of cattle type I interferon genes for 4 different subtypes of interferon have been ordered. (IFN-aA, IFN-aC, IFN-β, IFN-τ) with the addition of restriction sites and a hexahistidine (6×His) tag. To design the primers, the target gene was divided into overlapping oligonucleotides with a length of 60 bp with an overlap of 16-20 bp at an annealing temperature of 50 °C ± 3 °C.
Four cattle interferon genes of various subtypes were cloned into the pGEM-T plasmid vector for subsequent sequencing and confirmation of the correctness of their sequences. Cloning was performed using the pGEM-T Easy Vector System (Promega) in accordance with the manufacturer’s instructions.
An SSG-P-300-C preparative medium-pressure chromatograph with Sepure Protein Purification System in a custom configuration was purchased.
Genetically engineered structures have been created for the protein expression of 4 subtypes of cattle interferon in the bacterial expression system. Previously sequenced, verified clones of cattle interferon genes cloned into the pGEM-T vector were used to obtain expression constructs. Target fragments were excised from recombinant plasmids pGEM-T::IFN-aA, pGEM-T::IFN-aC, pGEM-T::IFN-β and pGEM-T::IFN-τ using appropriate restriction enzymes flanking the insertion region.