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[Prokaryotic expression and immunization of glutamyl tRNA synthetase of Streptococcus suis].
- Source :
-
Wei sheng wu xue bao = Acta microbiologica Sinica [Wei Sheng Wu Xue Bao] 2010 Mar; Vol. 50 (3), pp. 418-22. - Publication Year :
- 2010
-
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.<br />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).<br />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.
- Subjects :
- Amino Acid Sequence
Animals
Bacterial Vaccines genetics
Blotting, Western
Escherichia coli genetics
Female
Freund's Adjuvant immunology
Glutamate-tRNA Ligase chemistry
Glutamate-tRNA Ligase genetics
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Random Allocation
Sequence Homology, Amino Acid
Streptococcus suis genetics
Bacterial Vaccines immunology
Bacterial Vaccines metabolism
Escherichia coli metabolism
Glutamate-tRNA Ligase immunology
Glutamate-tRNA Ligase metabolism
Streptococcus suis enzymology
Subjects
Details
- Language :
- Chinese
- ISSN :
- 0001-6209
- Volume :
- 50
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Wei sheng wu xue bao = Acta microbiologica Sinica
- Publication Type :
- Academic Journal
- Accession number :
- 20499650