High level soluble expression, one-step purification and characterization of HIV-1 p24 protein

Virol J. 2011 Jun 22:8:316. doi: 10.1186/1743-422X-8-316.

Abstract

Background: P24 protein is the major core protein of HIV virus particle and has been suggested as a specific target for antiviral strategies. Recombinant p24 protein with natural antigenic activity would be useful for various studies, such as diagnostic reagents and multi-component HIV vaccine development. The aim of this study was to express and purify the p24 protein in soluble form in E.coli.

Results: According to the sequence of the p24 gene, a pair of primers was designed, and the target sequence of 700 bp was amplified using PCR. The PCR product was cloned into pQE30 vector, generating the recombinant plasmid pQE30-p24. SDS-PAGE analysis showed that the His-tagged recombinant p24 protein was highly expressed in soluble form after induction in E. coli strain BL21. The recombinant protein was purified by nickel affinity chromatography and used to react with HIV infected sera. The results showed that the recombinant p24 protein could specifically react with the HIV infected sera. To study the immunogenicity of this soluble recombinant p24 protein, it was used to immunize mice for the preparation of polyclonal antibody. Subsequent ELISA and Western-Blot analysis demonstrated that the p24 protein had proper immunogenicity in inducing mice to produce HIV p24 specific antibodies.

Conclusion: In this work, we report the high level soluble expression of HIV-1 p24 protein in E. coli. This soluble recombinant p24 protein specifically react with HIV infected sera and elicit HIV p24 specific antibodies in mice, indicating this soluble recombinant p24 protein could be a promising reagent for HIV diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / biosynthesis*
  • Antigens, Viral / genetics
  • Antigens, Viral / immunology
  • Antigens, Viral / isolation & purification*
  • Chromatography, Affinity / methods
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Female
  • Gene Expression*
  • HIV Antibodies / blood
  • HIV Antibodies / immunology
  • HIV Core Protein p24 / biosynthesis*
  • HIV Core Protein p24 / genetics
  • HIV Core Protein p24 / immunology
  • HIV Core Protein p24 / isolation & purification*
  • Mice
  • Mice, Inbred BALB C
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification

Substances

  • Antigens, Viral
  • HIV Antibodies
  • HIV Core Protein p24
  • Recombinant Proteins
  • p24 protein, Human Immunodeficiency Virus Type 1