Expression of the recombinant C-terminal of the S1 domain and N-terminal of the S2 domain of the spike protein of porcine epidemic diarrhea virus-Reference-Cited by-同舟云学术

Expression of the recombinant C-terminal of the S1 domain and N-terminal of the S2 domain of the spike protein of porcine epidemic diarrhea virus

Published:2021-11-12 Issue: Volume: Page:2913-2918
ISSN:2231-0916
Container-title:Veterinary World
language:en
Short-container-title:Vet World

Author:

Sritun Jiraporn¹,Inthong Natnaree²^ORCID,Jala Siriluk³,Phatthanakunanan Sakuna³,Satchasataporn Khomson²^ORCID,Sirinarumitr Kaitkanoke⁴^ORCID,Lertwatcharasarakul Preeda⁵,Sirinarumitr Theerapol⁵

Affiliation:

1. Bio-Veterinary Sciences Program, Faculty of Veterinary Medicine, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.

2. Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.

3. Kamphaeng Saen Veterinary Diagnosis Center, Faculty of Veterinary Medicine, Kasetsart University, Nakhon Pathom, 73140, Thailand.

4. Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.

5. Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.

Abstract

Background and Aim: Porcine epidemic diarrhea virus (PEDV) causes severe diarrhea in suckling piglets, leading to severe economic losses in the swine industry. Commercial vaccines have limited effectiveness against different genogroups of PEDV and the shedding of virus. The C-terminal of the S1 domain and the N-terminal of the S2 domain (S1-2) protein of the spike (S) protein have four neutralizing epitopes. However, research on the expression of the S1-2 segment of the S gene has been limited. In this study, we expressed a recombinant S1-2 protein of the S protein of the PEDV Thai isolate and characterized the immunological properties of the recombinant S1-2 protein. Materials and Methods: The S1-2 segment of the S gene of the PEDV Thai isolate (G2b) was amplified, cloned into the pBAD202/D-TOPO® vector (Invitrogen, Carlsbad, CA, USA), and expressed in Escherichia coli. The optimum concentration of arabinose and the optimum induction time for the expression of the recombinant S1-2 protein were determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The immunogenic reactivity of the recombinant S1-2 protein was determined using Western blot analysis with rabbit polyclonal antibodies against the SM98 strain of PEDV (G1a). Results: The recombinant S1-2 segment of the S gene of the PEDV Thai isolate protein was cloned and the recombinant S1-2 protein was successfully expressed. The optimum concentration of arabinose and the optimum induction time for the induction of the recombinant S1-2 protein were 0.2% and 8 h, respectively. The recombinant S1-2 protein reacted specifically with both rabbit anti-histidine polyclonal antibodies and rabbit anti-PEDV polyclonal antibodies. Conclusion: The recombinant S1-2 protein reacted with rabbit anti-PEDV polyclonal antibodies induced by the different PEDV genogroup. Therefore, the recombinant S1-2 protein may be a useful tool for the development of a diagnostic test for PEDV or for a vaccine against PEDV.

Funder

Kasetsart University

Publisher

Veterinary World

Subject

General Veterinary

Link

http://www.veterinaryworld.org/Vol.14/November-2021/10.pdf

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