Removal of 3C protease from the 3ABC improves expression, solubility, and purification of the recombinant 3AB of foot-and-mouth disease virus.
3C removal
Bacterial gene expression
FMDV
Immunoreactivity
Insolubility
Non-structural 3ABC
Recombinant 3AB
Journal
Virus genes
ISSN: 1572-994X
Titre abrégé: Virus Genes
Pays: United States
ID NLM: 8803967
Informations de publication
Date de publication:
Feb 2021
Feb 2021
Historique:
received:
02
08
2020
accepted:
21
11
2020
pubmed:
6
1
2021
medline:
26
8
2021
entrez:
5
1
2021
Statut:
ppublish
Résumé
During an ongoing outbreak of Foot-and-Mouth Disease Virus (FMDV), it is crucial to distinguish naturally infected from vaccinated seropositive animals. This would support clinical assessment and punctual vigilance. Assays based on 3ABC non-structural protein as an antigen are reliable for this intention. However, the insolubility and degradation of recombinant 3ABC during expression and purification are serious challenges. In this study, alternatively to expressing the recombinant 3ABC (r3ABC), we expressed the 3AB coding sequence (~672 bp) as a recombinant protein (r3AB) with a molecular mass of ~26 KDa. Analytical data from three-dimensional structure, hydrophilicity, and antigenic properties for 3ABC and 3AB exhibited the 3C protein as a hydrophobic, while 3AB as a hydrophilic and highly antigenic protein. The expressed r3AB was recovered as a completely soluble matter after merely native purification, unlike the full expressed r3ABC. Immunoreactivity of r3AB to anti-FMDV antibody in infected sera with different FMDV serotypes was confirmed by the western blot and indirect ELISA. Besides, the authentic antigenicity of purified r3AB was demonstrated through its ability to induce specific seroconversion in mice. Summarily, the removal of 3C: has influenced neither 3D structure nor antigenic properties of the purified r3AB, overcame insolubility and degradation of the r3ABC, and generated a potential superior antigen (r3AB) for herd screening of animals to any FMDV serotype.
Identifiants
pubmed: 33400101
doi: 10.1007/s11262-020-01815-8
pii: 10.1007/s11262-020-01815-8
doi:
Substances chimiques
Recombinant Proteins
0
Viral Nonstructural Proteins
0
3C Viral Proteases
EC 3.4.22.28
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
72-82Subventions
Organisme : Academy of Scientific Research and Technology
ID : National Strategy Programs for Biotechnology and Genetic Engineering (ID: 31ح/2018)
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