Cloning and expression of toxoplasma gondii dense granule antigen 2 (GRA2) gene by Pichia Pastoris

Lau, Y.L. and Fong, M.Y. and Idris, M.M. and Ching, X.T. (2012) Cloning and expression of toxoplasma gondii dense granule antigen 2 (GRA2) gene by Pichia Pastoris. Southeast Asian Journal of Tropical Medicine and Public Health, 43 (1). pp. 10-16. ISSN 0125-1562

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Abstract

Detection of Toxoplasma gondii infection is essential in pregnant women and immunosuppressed patients. Numerous studies have shown that the recombinant production of several Toxoplasma antigens, including dense granule antigens (GRAs) has high potential as diagnostic reagents. In the present study, we produced GRA2 using Pichia pastoris system. RNA of T. gondii RH strain tachyzoite was used as a template to produce cDNA clones of full-length GRA2 via reverse transcriptase PCR. Amplicons were inserted into pPICZ alpha A and the recombinant plasmid transformed into P. pastoris, X-33 strain. The expressed recombinant protein was identified by SDS-PAGE and Western blotting. A recombinant protein of similar to 28 kDa was produced, which could be detected by toxoplasmosis positive human sera indicating that the recombinant protein retained its antigenicity. The present study indicates that P. pastoris-expressed GRA2 should be useful for detection of Toxoplasma infection.

Item Type: Article
Additional Information: Correspondence: Dr Lau Yee Ling, Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Tel: 6 03 7967 4749; Fax: 6 03 7967 4754 E-mail: lauyeeling@um.edu.my
Uncontrolled Keywords: Toxoplasma gondii, GRA2, expression, Western blot
Subjects: R Medicine
Divisions: Faculty of Medicine
Depositing User: Fong Mun Yik
Date Deposited: 05 Jun 2012 02:03
Last Modified: 05 Jun 2012 02:03
URI: http://eprints.um.edu.my/id/eprint/3206

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