Codon Optimization, Expression in Escherichia coli, and Immunogenicity of Recombinant Chinese Sacbrood Virus (CSBV) Structural Proteins VP1, VP2, and VP3

PLoS One. 2015 Jun 12;10(6):e0128486. doi: 10.1371/journal.pone.0128486. eCollection 2015.

Abstract

Chinese sacbrood virus (CSBV) is a small RNA virus family belonging to the genus Iflavirus that causes larval death, and even the collapse of entire bee colonies. The virus particle is spherical, non-enveloped, and its viral capsid is composed of four proteins, although the functions of the structural proteins are unclear. In this study, we used codon recoding to express the recombinant proteins VP1, VP2, and VP3 in Escherichia coli. SDS-PAGE analysis and Western blotting revealed that the target genes were expressed at high levels. Mice were then immunized with the purified, recombinant proteins, and antibody levels and lymphocyte proliferation were analyzed by ELISA and the MTT assay, respectively. The results show that the recombinant proteins induced high antibody levels and promoted lymphocyte proliferation. Polyclonal antibodies directed against these proteins will aid future studies of the molecular pathogenesis of CSBV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / blood
  • Antibodies, Viral / immunology
  • Antigens, Viral / immunology
  • Capsid Proteins / genetics*
  • Capsid Proteins / immunology*
  • Codon*
  • Escherichia coli / genetics*
  • Gene Expression*
  • Immunization
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Lymphocyte Activation / immunology
  • Mice
  • Plasmids / genetics
  • RNA Viruses / genetics*
  • RNA Viruses / immunology
  • Recombinant Proteins*

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Capsid Proteins
  • Codon
  • Immunoglobulin G
  • Recombinant Proteins

Associated data

  • figshare/10.6084/M9.FIGSHARE

Grants and funding

This work was supported by grants from the National Natural Science Foundation of China (No. 31372435) and the Science and Technology Department of Liaoning Province (No. 2014022047). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.