EXPRESSION OF THE SURFACE ANTIGEN BRUCELLA ABORTUS OMR16 IN NICOTIANA BENTHAMIANA PLANT
Authors:   Doctor of Veterinary Sciences, Professor, Department of Biological Safety, Faculty of Veterinary Medicine, KazNAU, Almaty, Kazakhstan, Ilgekbaeva, G.D. ,   https://orcid.org/0000-0001-7140-2961, Makhashov E.Sh. ,   сandidate of Veterinary Sciences, Acting Professor, Department of Biological Safety, Faculty of Veterinary Medicine, KazNAU, Almaty, Kazakhstan; https://orcid.org/0000-0003-1922-7144, Tulepova G. ,   doctoral student, Department of Biological Safety, Faculty of Veterinary Medicine, KazNAU, Almaty, Kazakhstan; https://orcid.org/0000-0002-0681-3788, Yessimkhankyzy D. ,   master student, Department of Biological Safety, Faculty of Veterinary Medicine, KazNAU, Almaty, Kazakhstan; https://orcid.org/0000-0001-7035-913X, Sadiev S.T. ,   director of the Center for Applied Agricultural Research, KazNAU, Almaty, Kazakhstan; sadiev15@mail.ru; https://orcid.org/0000-0003-1470-2612
Published: October 2020

Brucellosis is one of the most contagious and infectious diseases with high incidence rates of cattle and humans in Kazakhstan. Using modern biotechnology techniques to develop vaccines that are reliable and affordable for farmers is an alternative solution to the problem. Plant viruses are often used as a vector for obtaining the expression of antigens of the pathogen. The grape virus A (BAB) is widely used among plant viruses. Brucella membrane proteins are the main objects of this research for futher development of vaccines or diagnostic texts against brucellosis, Membrane proteins (OMPs) are cell specific surface antigens that are immunogenic. OMPs are ideal candidates for the production of recombinant brucellosis vaccines. The object of the study was the outer membrane protein (Omp16), which plays an important role in the suppression of TNF-α production in macrophages. In this study, molecular cloning and analysis of the expression of the Omp16 gene, which was used to express the recombinant protein in plants, was carried out. We selected brucella from the vaccine strain of Brucella abortus 19, and the plant Nicotiana benthamiana, as the subjects for our research, since they widely used for the production of recombinant proteins, and they both appropriate for molecular genetic research. A viral vector was constructed to express the brucellosis antigen Omp16 in Nicotiana benthamiana plants. Nineteen explants were used for the regeneration of transgenic plants. As a result of this studies, the introduced gene of Omp16 was under the subgenomic promoter control of the ORF4 and was successfully expressed while maintaining the efficiency of expression in transgenic plants. The efficiency of viral vectors was evaluated at the level of transcription during expression of the protein Omp16 with viral proteins. The entire leaf blade was infiltrated; the density of Agrobacteria was 0.7. We were able to obtained transgenic plants Nicotiana benthamiana carrying the gene of capsid protein BAB, and the expression of the membrane antigen Omp16 in the viral vector was achieved by replacing the ORF4 with the Omp16 gene. The development of transgenic plants was carried out using agrobacterial transformation.