Expression of a Recombinant Protein Which Can Be Recognized by Antibody to Toxoplasma gondii SAG1
Journal
Bio Formosa
Journal Volume
43
Journal Issue
1
Pages
1-10
Date Issued
2008
Author(s)
Lin,D. S.
Abstract
Toxoplasma gondii (T. gondii) infection can cause serious consequences in both humans and animals. Toxoplasmosis is a widespread zoonotic disease and thus is of economic as well as public health importance. Because toxoplasmosis lacks specific clinical signs or syndromes, the application of diagnostic techniques is required to detect the infection. Detection of antibodies to T. gondii has been very useful as an indicator of infection. Nevertheless, no final diagnosis will be made unless a native 30-kilodalton T. gondii-specific surface antigen 1 (SAG1) is recognized by the test samples. SAG1 is highly conserved among various strains of T. gondii and thus is a very useful molecule for diagnosis of toxoplasmosis. In this study, SAG1 gene was amplified and further purified as a 1,011-base pair nucleotide which was then ligated with pET-24b vector as a 6,320-base pair recombinant pET-24b/SAG1. After transformation, a 63-kilodalton protein was expressed in calcium chloride-treated competent Escherichia coli. This recombinant protein could be identified by T. gondii-infected cat sera which also were able to recognize SAG1 molecule by immunoblotting. Furthermore, all 30 sera which could recognize expressed recombinant protein were also positive in kinetics-based enzyme linked immunosorbent assay (ELISA) with recombinant protein. Therefore, ELISA with this expressed recombinant protein, combining the advantages of both traditional ELISA and time consuming immunoblotting, will be a powerful tool for surveying a large number of serum samples with excellent sensitivity and specificity. The potential candidate of this recombinant protein for vaccine development is also discussed.
SDGs
Type
journal article