[Cloning, fusion expression and identification of thioredoxin encoding gene from Toxoplasma gondii].

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.