Advances in Animal and Veterinary Sciences

Research Article
Adv. Anim. Vet. Sci. 9(4): 581-587
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Boonyasit Porngarm1, Aktsar Roskiana Ahmad1, Kate Neelasawee3, Pichai Joiphaeng3, Tawatchai Hoonsuwan3, Kaewta Rattanapisit1,2, Balamurugan Shanmugaraj1,2*, Waranyoo Phoolcharoen1,2*

1Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand; 2Research unit for Plant-produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand; 3B.F. Feed Company Limited, Thung Song Hong, Lak Si, Bangkok, Thailand.

Abstract | Porcine reproductive and respiratory syndrome virus (PRRSV), the causative agent of Porcine reproductive and respiratory syndrome (PRRS) in pigs, is an enveloped positive strand RNA virus belongs to the family Arteriviridae. PRRSV poses severe threat to swine industry worldwide. Routine surveillance, rapid timely diagnosis of PRRSV infection is crucial for effective monitoring and disease control. Serological tests based on highly conserved viral nucleocapsid (N) protein are widely used for the detection of antibodies to PRRSV. Here, we investigated the potential of plant system to produce PRRSV N-protein in order to use as a reagent for the diagnosis of PRRSV infection. In this regard, the codon-optimised PRRSV N-protein coding sequence was cloned in geminiviral vector and expressed in Nicotiana benthamiana. The transient expression conditions such as effective gene construct, leaf harvesting time and Agrobacterium cell density were optimized for maximal protein production. Further, by using western blot assay, with a plant-produced protein we sought to detect the antibodies against PRRSV in pig sera. Preliminary testing revealed that the recombinant protein was identified by antibodies present in pig sera. However sensitivity/specificity and correlation of results with other assays needs to be evaluated. This proof of concept study indicated that plant-produced N-protein can likely be used as diagnostic reagent and demonstrate the potential of plant expression system for the production of recombinant PRRSV antigens.

Keywords | Nicotiana benthamiana; Nucleocapsid protein; Plant-produced protein; Porcine reproductive and respiratory syndrome virus; Transient expression