CLONING AND EXPRESSION OF P30 GENE FRAGMENT OF TOXOPLASMA GONDII

Objective To construct a recombinant plasmid containing P30 gene of Toxoplasma gondii and obtain recombinant protein Methods P30 gene fragment was amplified from genomic DNA of T. gondii RH strain by PCR. The purified PCR fragment was ligated into the sites of pET-30a(+). The recombinant vector was transformed into Escherichia coli DH5α. The positive clones were screened and identified by PCR from bacteria directly and digested via double enzymes cut. Sequencing of recombinant plasmid was performed. The recombinant plasmid was transformed into E. coli BL21(DE3), the expression of protein was induced by IPTG. The expression product was identified by SDS-PAGE and Western blot which was proceeded with P30 McAb produced by our laboratory. Results The size of amplified gene was 750 bp, the molecular weight of the expression protein was 30 ku. The expressed protein was futher identified by Western blot which was performed with P30 McAb. Conclusion The recombinant plasmid was successfully constructed, the protein of P30 gene of T. gondii over-expressed laid solid foundations for the diagnosis of toxoplasmosis as well as study of vaccines.