RESUMO
OBJECTIVE: To clone and express the thioredoxin (Trx) from RH strain tachyzoites of Toxoplasma gondii, establish the prokaryotic expression vector and purify the recombinant protein, then produce the polyclonal anti-Trx antibody in rabbits. METHODS: Trx fragment was amplified by PCR and cloned into the pET-28a (+) vector, and the recombinant protein was induced with IPTG and purified by Ni-NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. RESULTS: The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET-28a (+) was usefully constructed, and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also obtained. The specific band of TRX was detected by Western blotting. CONCLUSIONS: Western blotting can detect the specificity of polyclonal anti-Trx antibody, which will facilitate the biological functions of Trx.
