Salmonella flagellin acted as an effective fusion partner for expression of Plasmodium falciparum surface protein 25 in Escherichia coli

Hum Vaccin Immunother. 2016 Sep;12(9):2362-4. doi: 10.1080/21645515.2016.1171443. Epub 2016 Apr 8.

Abstract

Plasmodium falciparum surface protein 25 (Pfs25) is a hard-to-express and hard-to-solubilize protein in Escherichia coli. To overcome this problem, the phase 1 flagellin of Salmonella enterica serovar Typhimurium (FliC) was used as a fusion partner for Pfs25. The fusion expression of Pfs25 with FliC greatly enhanced the expression level and solubility of Pfs25 in E. coli BL21(DE3). The Ni-purified fusion protein of FliC-Pfs25 was recognized by two anti-Pfs25 monoclonal antibodies. By comparison, it was shown that the Pfs25 within FliC-Pfs25 contained epitopes similar or identical to those on Pichia pastoris-produced Pfs25. The data obtained from this study demonstrated that the fusion with Salmonella flagellin greatly improved the expression of Pfs25 in E. coli.

Keywords: Escherichia coli; Fusion partner; Pfs25; Plasmodium falciparum; Salmonella flagellin.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Protozoan / immunology
  • Epitopes / immunology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Flagellin / biosynthesis*
  • Flagellin / genetics
  • Gene Expression*
  • Mice, Inbred BALB C
  • Plasmodium falciparum / genetics
  • Plasmodium falciparum / immunology*
  • Protozoan Proteins / biosynthesis*
  • Protozoan Proteins / genetics
  • Protozoan Proteins / immunology
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Salmonella typhimurium / genetics
  • Salmonella typhimurium / immunology*

Substances

  • Antibodies, Monoclonal
  • Antibodies, Protozoan
  • Epitopes
  • Pfs25 protein, Plasmodium falciparum
  • Protozoan Proteins
  • Recombinant Fusion Proteins
  • Flagellin