Thiruvengadam, G.; Init, I.; Fong, M.Y.; Lau, Y.L. (2011) Optimization of the expression of surface antigen SAG1/2 of Toxoplasma gondii in the yeast Pichia pastoris. Tropical Biomedicine, 28 (3). pp. 506-513. ISSN 0127-5720
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Abstract
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 30 masculineC, 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.
| Item Type: | Article |
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| Creators: |
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| Journal or Publication Title: | Tropical Biomedicine |
| Additional Information: | Thiruvengadam, G Init, I Fong, M Y Lau, Y L eng Evaluation Studies Research Support, Non-U.S. Gov't Malaysia 2012/03/22 06:00 Trop Biomed. 2011 Dec;28(3):506-13. |
| Uncontrolled Keywords: | Ammonium Sulfate/chemistry Antigens, Protozoan/*biosynthesis/diagnostic use/genetics/isolation & purification Carbon/metabolism Chromatography, Affinity Cloning, Molecular Culture Media/chemistry Dialysis Fractional Precipitation *Gene Expression Hydrogen-Ion Concentration Methanol/metabolism Pichia/*genetics Protozoan Proteins/*biosynthesis/diagnostic use/genetics/isolation & purification Recombinant Fusion Proteins/biosynthesis/diagnostic use/genetics/isolation & purification Sensitivity and Specificity Serologic Tests/methods Temperature Toxoplasmosis/diagnosis |
| Subjects: | R Medicine |
| Divisions: | Faculty of Medicine |
| Depositing User: | Mr Zainal Sabri |
| Date Deposited: | 06 Dec 2012 14:57 |
| Last Modified: | 06 Dec 2012 14:57 |
| URI: | http://eprints.um.edu.my/id/eprint/4076 |
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