Co-expression of self-cleaved multiple proteins derived from Porcine Reproductive and Respiratory Syndrome Virus by bi-cistronic and tri-cistronic DNA vaccines

Journal article

Authors/Editors

Strategic Research Themes

Publication Details

Author list: Meas S., Mekvichitsaeng P., Roshorm Y.M.

Publisher: Elsevier

Publication year: 2021

Journal: Protein Expression and Purification (1046-5928)

Volume number: 177

ISSN: 1046-5928

URL: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85091592667&doi=10.1016%2fj.pep.2020.105763&partnerID=40&md5=b602a95036ed4f327bf3f5407bc624b6

Languages: English-Great Britain (EN-GB)

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Abstract

Porcine Reproductive and Respiratory Syndrome caused by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) remains one of the important diseases in swine industry. A vaccine that is safe, effective and also elicit broad immune response against multiple antigens is desirable. In this study, we developed multi-cistronic DNA vaccines capable of co-expressing multiple structural proteins derived from PRRSV. To preserve the structure and function of each antigen protein, we employed self-cleaving 2A peptides to mediate separation of multiple proteins expressed by multi-cistronic genes. Six bi-cistronic genes encoding PRRSV GP5 and M proteins were generated, by which each construct contains different 2A sequences derived from Foot-and-mouth disease virus (F2A), porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A) either with or without furin cleavage site (Fu). Vectored by the mammalian expression plasmid pTH, all six bi-cistronic genes co-expressed the proteins GP5 and M at comparable level. Importantly, all six types of 2A sequences could mediate a complete self-cleavage of the GP5 and M. We next generated tri-cistronic DNA vaccines co-expressing the PRRSV proteins GP5, M and N. All homologous and heterologous combinations of P2A and F2A in tri-cistronic genes yielded a complete self-cleavage of the GP5, M and N proteins. Our study reports a success in co-expression of multiple PRRSV structural proteins in discrete form from a single vaccine and confirms feasibility of developing one single vaccine that provides broad immune responses against PRRSV. © 2020 Elsevier Inc.

Keywords

2A peptide, Furin cleavage, Multi-cistronic, PRRSV, Self-cleaving peptide