Back to Search
Start Over
High Expression of Water-Soluble Recombinant Antigenic Domains ofToxoplasma gondii Secretory Organelles
- Source :
- The Korean Journal of Parasitology
- Publication Year :
- 2014
- Publisher :
- Korean Society for Parasitology, 2014.
-
Abstract
- Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with GRA225-105, GRA339-138, ROP2324-561, and MIC21-284 domains had respectively higher value of IgG avidity. The rGST-GRA225-105 and rGST-GRA339-138 were soluble, while rGST-ROP2324-561 and rGST-MIC21-284 were not. GRA231-71, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The rGST-GRA231-71-ROP2324-561, a chimeric protein, appeared to be soluble. Moreover, rGST-GRA231-71-MIC21-284 was also soluble and had higher IgG avidity comparing to rGST-MIC21-284. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.
- Subjects :
- MIC2
Antibody Affinity
Antibodies, Protozoan
Gene Expression
Toxoplasma gondii
intrinsically unstructured domain
Antigens, Protozoan
Immunoglobulin G
law.invention
ROP2
Plasmid
Antigen
law
Gene expression
Animals
Serologic Tests
Mice, Inbred BALB C
biology
antigenic domain
bioinformatics
biology.organism_classification
Molecular biology
Fusion protein
Recombinant Proteins
Infectious Diseases
Secretory protein
Solubility
GRA3
GRA2
IgG avidity
biology.protein
Recombinant DNA
Original Article
Parasitology
Toxoplasma
Toxoplasmosis
Subjects
Details
- ISSN :
- 17380006
- Volume :
- 52
- Database :
- OpenAIRE
- Journal :
- The Korean Journal of Parasitology
- Accession number :
- edsair.doi.dedup.....c1820c04330aba2352830c844d831269
- Full Text :
- https://doi.org/10.3347/kjp.2014.52.4.367