title: High level soluble expression, one-step purification and characterization of HIV-1 p24 protein.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Baozhong
authors.initials: B
authors.lastname: Zhang
authors.forename: Dabin
authors.initials: D
authors.lastname: Liu
authors.forename: Zuoyi
authors.initials: Z
authors.lastname: Bao
authors.forename: Bin
authors.initials: B
authors.lastname: Chen
authors.forename: Cun
authors.initials: C
authors.lastname: Li
authors.forename: Huanhuan
authors.initials: H
authors.lastname: Jiang
authors.forename: Xiaona
authors.initials: X
authors.lastname: Wang
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Mi
authors.forename: Xiaoping
authors.initials: X
authors.lastname: An
authors.forename: Jun
authors.initials: J
authors.lastname: Lu
authors.forename: Yigang
authors.initials: Y
authors.lastname: Tong
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D015483:HIV Antibodies / Q000097:blood / Q000276:immunology; D016655:HIV Core Protein p24 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000956:Antigens, Viral; D015483:HIV Antibodies; D016655:HIV Core Protein p24; D011994:Recombinant Proteins; C514799:p24 protein, Human Immunodeficiency Virus Type 1