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

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 doma...

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Autores principales:	Jiraporn Sritun, Natnaree Inthong, Siriluk Jala, Sakuna Phatthanakunanan, Khomson Satchasataporn, Kaitkanoke Sirinarumitr, Preeda Lertwatcharasarakul, Theerapol Sirinarumitr
Formato:	article
Lenguaje:	EN
Publicado:	Veterinary World 2021
Materias:	diagnostics genogroup porcine epidemic diarrhea virus recombinant protein vaccine Animal culture SF1-1100 Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/83d8f349b5b24c80889c660ae1531adf
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spelling	oai:doaj.org-article:83d8f349b5b24c80889c660ae1531adf2021-11-12T11:42:33ZExpression of the recombinant C-terminal of the S1 domain and N-terminal of the S2 domain of the spike protein of porcine epidemic diarrhea virus10.14202/vetworld.2021.2913-29180972-89882231-0916https://doaj.org/article/83d8f349b5b24c80889c660ae1531adf2021-11-01T00:00:00Zhttp://www.veterinaryworld.org/Vol.14/November-2021/10.pdfhttps://doaj.org/toc/0972-8988https://doaj.org/toc/2231-0916Background 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.Jiraporn SritunNatnaree InthongSiriluk JalaSakuna PhatthanakunananKhomson SatchasatapornKaitkanoke SirinarumitrPreeda LertwatcharasarakulTheerapol SirinarumitrVeterinary Worldarticlediagnosticsgenogroupporcine epidemic diarrhea virusrecombinant proteinvaccineAnimal cultureSF1-1100Veterinary medicineSF600-1100ENVeterinary World, Vol 14, Iss 11, Pp 2913-2918 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	diagnostics genogroup porcine epidemic diarrhea virus recombinant protein vaccine Animal culture SF1-1100 Veterinary medicine SF600-1100
spellingShingle	diagnostics genogroup porcine epidemic diarrhea virus recombinant protein vaccine Animal culture SF1-1100 Veterinary medicine SF600-1100 Jiraporn Sritun Natnaree Inthong Siriluk Jala Sakuna Phatthanakunanan Khomson Satchasataporn Kaitkanoke Sirinarumitr Preeda Lertwatcharasarakul Theerapol Sirinarumitr 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
description	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.
format	article
author	Jiraporn Sritun Natnaree Inthong Siriluk Jala Sakuna Phatthanakunanan Khomson Satchasataporn Kaitkanoke Sirinarumitr Preeda Lertwatcharasarakul Theerapol Sirinarumitr
author_facet	Jiraporn Sritun Natnaree Inthong Siriluk Jala Sakuna Phatthanakunanan Khomson Satchasataporn Kaitkanoke Sirinarumitr Preeda Lertwatcharasarakul Theerapol Sirinarumitr
author_sort	Jiraporn Sritun
title	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
title_short	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
title_full	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
title_fullStr	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
title_full_unstemmed	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
title_sort	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
publisher	Veterinary World
publishDate	2021
url	https://doaj.org/article/83d8f349b5b24c80889c660ae1531adf
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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

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