Optimization of the expression of surface antigen SAG1/2 of Toxoplasma gondii in the yeast Pichia pastoris
Surface antigens are the most abundant proteins found on the surface of the parasite Toxoplasma gondii. Surface antigen 1 (SAG1) and Surface antigen 2 (SAG2) remain the most important and extensively studied surface proteins. These antigens have been identified to play a role in host cell invasion,...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | en |
| Published: |
Malaysian Society of Parasitology and Tropical Medicine
2011
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| Subjects: | |
| Online Access: | http://eprints.um.edu.my/3219/1/Optimization_of_the_expression_of_surface_antigent_SAG1.pdf http://eprints.um.edu.my/3219/ https://drive.google.com/file/d/0B75lcx0mfp2OWXQtSUFDcDZ2UXM/view |
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| Summary: | Surface antigens are the most abundant proteins found on the surface of the parasite Toxoplasma gondii. Surface antigen 1 (SAG1) and Surface antigen 2 (SAG2) remain the most important and extensively studied surface proteins. These antigens have been identified to play a role in host cell invasion, immune modulation, virulence attenuation. Recombinant SAG1/2 was cloned and expressed in yeast Pichia pastoris. We describe here optimization of critical parameters involved in high yield expression of the recombinant SAG1/2. Our results suggest that recombinant SAG1/2 were best expressed at 30ºC, pH 6 and 1 methanol as the carbon source by X33 Pichia cells. Additional optimizations included the downstream process such as ammonium sulphate precipitation and dialysis. The fusion protein was purified using Ni-NTA purification system with 80 recovery. The purified protein was 100 specific and sensitive in detection of toxoplasmosis. |
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