Abstract
ObjectiveA 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.ResultsTwo 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.ConclusionsThese results demonstrate that chloroplasts are a suitable system for the production of a candidate subunit vaccine against sheep pox.
Highlights
Sheep pox is a highly contagious disease of small ruminants, which has a wide global distribution area
Plants represent an attractive system for recombinant vaccine production because of their ease of scalability, low cost of production compared to other eukaryotic systems such as yeast and animal cells, and the absence of human and animal pathogens in the production host (Fischer and Buyel 2020)
The analysis showed the presence of four potential sites for O-linked glycosylation (Ser6, Thr15, Thr26, and Ser27) in SPPV117
Summary
Sheep pox is a highly contagious disease of small ruminants, which has a wide global distribution area It causes large economic losses in livestock animals; it is classified as a notifiable disease by the World Organization for Animal Health Producing recombinant proteins in plant chloroplasts has several advantages over nuclear integration of the transgene including an absence of gene silencing and position effects. Another advantage is a high expression level of a target gene. A number of antigens have been successfully produced in chloroplasts, including antigens of dengue virus, poliovirus, and Mycobacterium tuberculosis (van Eerde et al 2019; Daniell et al 2019; Saba et al 2019), providing the rationale for producing a plant-based, low-cost animal vaccine against sheep pox
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