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
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 30ÂºC, 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|
- Thiruvengadam, G.
- Init, I.
- Fong, M.Y.(Department of Parasitology, Faculty of Medicine Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA)
- Lau, Y.L.
|Journal or Publication Title: ||Tropical Biomedicine|
|Additional Information: ||Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia * Corresponding author email: firstname.lastname@example.org|
|Uncontrolled Keywords: ||Parasitology|
|Subjects: ||R Medicine|
|Divisions: ||Faculty of Medicine|
|Depositing User: ||Fong Mun Yik|
|Date Deposited: ||07 Jun 2012 15:06|
|Last Modified: ||07 Jun 2012 15:06|
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