The production of molecular clones of Toxoplasma gondii-derived heat shock protein 70 kDa

Toxoplasmosis is a common and widespread infection in humans and many other species of warm–blooded animals. Toxoplasma gondii-derived heat shock protein 70 (Hsp70) may play an important role in the virulence of Toxoplasma gondii (T. gondii). In the present study, T. gondii Hsp70 was amplified by polymerase chain reaction (PCR) from the DNA of the T. gondiitachyzoite RH strain through the use of specific primers with Xho1 and Xba1 restriction sites. The purified DNA fragment of the T. gondii Hsp70 gene was subcloned into the Xho1 and Xba1-digested eukaryotic expression vector, pcDNA3, and subsequently transformed into TOP10 chemically competent cells. A 2004 base pair (bp) band of PCR product was observed on the 0.8% agarose gel. The cDNA was inserted into the pTZ57R/T vector and then subcloned successfully into the pcDNA3 eukaryotic expression plasmid vector. The sequence of this amplified gene showed up to 100% homology with the target gene according to the Genbank database (Accession no. U82281).