Cloning and Molecular Characterization of the Recombinant CVB4E2 Immunogenic Viral Protein (rVP1), as a Potential Subunit Protein for Vaccine and Immunodiagnostic Reagent Candidate

Author:

Hadj Hassine Ikbel1,Gharbi Jawhar2ORCID,Amara Imene1,Alyami Ameera2,Subei Reem2,Almalki Mohammed2ORCID,Hober Didier3ORCID,M’hadheb Manel Ben1ORCID

Affiliation:

1. Virology and Antiviral Strategies Research Unit, Institute of Biotechnology, University of Monastir, BP74, Monastir 5000, Tunisia

2. Department of Biological Sciences, College of Science, King Faisal University, P.O. Box 380, Al-Ahsa 31982, Saudi Arabia

3. Laboratoire de Virologie ULR3610, Université de Lille et CHU de Lille, 59000 Lille, France

Abstract

The aim of the present study was, first, to clone the VP1 gene of the human coxsackievirus B4 strain E2 (CVB4E2) in the prokaryotic pUC19 plasmid expression vector then to compare it with the structural capsid proteins of the same strain using bioinformatic tools. PCR colony amplification followed through a restriction digestion analysis and sequencing process which affirmed the success of the cloning process. SDS-PAGE and Western Blotting were used to characterize the purified recombinant viral protein expressed in bacteria cells. The BLASTN tool revealed that the nucleotide sequence of the recombinant VP1 (rVP1) expressed by pUC19 highly matched the target nucleotide sequence of the diabetogenic CVB4E2 strain. Secondary structure and three-dimension structure prediction suggested that rVP1, such as wild-type VP1, is chiefly composed of random coils and a high percentage of exposed amino acids. Linear B-cell epitope prediction showed that several antigenic epitopes are likely present in rVP1 and CVB4E2 VP1 capsid protein. Additionally, phosphorylation site prediction revealed that both proteins may affect the signal transduction of host cells and can be involved in virus virulence. The present work highlights the usefulness of cloning and bioinformatics characterizations for gene investigation. Furthermore, the collected data are helpful for future experimental research related to the development of immunodiagnostic reagents and subunit vaccines based on the expression of immunogenic viral capsid proteins.

Funder

King Faisal University

Ministry of Higher Education and Scientific Research

Publisher

MDPI AG

Subject

Virology,Microbiology (medical),Microbiology

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