Production of the sheep pox virus structural protein SPPV117 in tobacco chloroplasts
Stanbekova G. Beisenov D. Nizkorodova A. Iskakov B. Warzecha H.
July 2021Springer Science and Business Media B.V.
Biotechnology Letters
2021#43Issue 71475 - 1485 pp.
Objective: A chloroplast transgenic approach was assessed in order to produce a structural protein SPPV117 of sheep pox virus in Nicotiana tabacum for the future development of a plant-based subunit vaccine against sheep pox. Results: Two DNA constructs containing SPPV117 coding sequence under the control of chloroplast promoter and terminator of psbA gene or rrn promoter and rbcL terminator were designed and inserted into the chloroplast genome by a biolistic method. The transgenic plants were selected via PCR analysis. Northern and Western blot analysis showed expression of the transgene at transcriptional and translational levels, respectively. The recombinant protein accumulated to about 0.3% and 0.9% of total soluble protein in leaves when expressed from psbA and rrn promoter, respectively. Plant-produced SPPV117 protein was purified using metal affinity chromatography and the protein yield was 19.67 ± 1.25 µg g−1 (FW). The serum of a sheep infected with the virus recognised the chloroplast-produced protein indicating that the protein retains its antigenic properties. Conclusions: These results demonstrate that chloroplasts are a suitable system for the production of a candidate subunit vaccine against sheep pox.
Sheep pox virus , SPPV117 protein , Transplastomic plants
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Protein and Nucleic Acids Research, M. Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan
Plant Biotechnology and Metabolic Engineering, Technical University of Darmstadt, Darmstadt, Germany
Protein and Nucleic Acids Research
Plant Biotechnology and Metabolic Engineering
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