[Prokaryotic expression and immunization of glutamyl tRNA synthetase of Streptococcus suis]

Wei Sheng Wu Xue Bao. 2010 Mar;50(3):418-22.
[Article in Chinese]

Abstract

Objective: Glutamyl tRNA Synthetase (Gts) is a protease which catalyzes esterification between tRNA and glutamine. The immunogenicity of Gts was evaluated through immunization and challenge experiment.

Methods and results: We cloned gts from the genome of SC19, and inserted it into prokaryotic expression plasmid pET28a-gts. The recombinant vector was transformed into E. coli BL21. Induced by IPTG, one 58 kD protein, was expressed and purified by using Ni - NTA column (Novagen). The purity of rGtS was 93.3% and the concentration of purified protein was 433 microg/mL. We proved the immunogenicity of recombinant protein rGtS by western blot analysis. We immunized Balb/c mice with rGtS and Freund's adjuvant, and after two boost vaccinations, all mice were challenged with 4 times LD50 amount of SC19 (1.2 x 10(9) CFU). The survive rate of vaccination group is 50% (4/8), significantly higher than blank vector control group (1/8).

Conclusion: These results proved that GtS has certain immunogenicity and can offer partial protection against high dose challenge. Therefore Gts could be a potential candidate of subunit vaccine against Streptococcus suis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / immunology*
  • Bacterial Vaccines / metabolism*
  • Blotting, Western
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Female
  • Freund's Adjuvant / immunology
  • Glutamate-tRNA Ligase / chemistry
  • Glutamate-tRNA Ligase / genetics
  • Glutamate-tRNA Ligase / immunology*
  • Glutamate-tRNA Ligase / metabolism*
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Random Allocation
  • Sequence Homology, Amino Acid
  • Streptococcus suis / enzymology*
  • Streptococcus suis / genetics

Substances

  • Bacterial Vaccines
  • Freund's Adjuvant
  • Glutamate-tRNA Ligase