Molecular cloning, expression and vaccination of a novel gene Sj-MA of Schistosoma japonicum

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Nov;35(11):981-5.

Abstract

In order to obtain new gene and to develop the new vaccine candidate of immunoprotection against Schistosoma japonicum (Sj), Sj adult worm cDNA library was screened with anti-sera to soluble male adult worm antigen and resulted in the discovery of a novel gene designated as Sj-MA. Sequence analysis showed that Sj-MA as a complete cDNA contains one open reading frame. It was deduced to contain 249 amino acid residues and encode a 28.8 kD soluble protein with plenty of phosphorylation sites, supposing its action in signal transduction. Furthermore, Sj-MA cDNA was cloned into a prokaryotic expression vector pGEX-5X to construct the recombinant plasmid which was transformed and highly expressed in E. coli as a 54.8 kD glutathione-S-transferase (GST) fusion protein. The fusion protein rSj-MA/GST could be recognized with both anti-male adult worm sera and anti-GST sera in Western blotting. Mice vaccinated with the fusion protein revealed significant worm reduction rate of 34.29% (P<0.001), compared with the control groups. Taken together, the novel gene Sj-MA can be expressed in E. coli as a fusion protein that can elicit immunity against Schistosoma japonicum, suggesting its potential as a new vaccine candidate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Helminth / blood
  • Antigens, Helminth / genetics*
  • Antigens, Helminth / immunology
  • Base Sequence
  • Blotting, Western
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Helminth Proteins / genetics*
  • Helminth Proteins / immunology
  • Helminth Proteins / metabolism
  • Immunoglobulin G / blood
  • Male
  • Mice
  • Molecular Sequence Data
  • Rabbits
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Schistosoma japonicum / genetics*
  • Schistosoma japonicum / immunology
  • Schistosomiasis mansoni / immunology
  • Schistosomiasis mansoni / prevention & control
  • Sequence Analysis, DNA
  • Vaccination

Substances

  • Antibodies, Helminth
  • Antigens, Helminth
  • DNA, Complementary
  • Helminth Proteins
  • Immunoglobulin G
  • Recombinant Fusion Proteins
  • Sj-MA protein, Schistosoma japonica
  • Glutathione Transferase

Associated data

  • GENBANK/AF519808