Expression and identification of P30 surface antigen from RH strain of Toxoplasma gondii / 中国血吸虫病防治杂志

ABSTRACT

Objective To express P30 surface antigen of RH strain of Toxoplasma gondii in E.coli BL21(DE3). Methods The P30 gene from Toxoplasma gondii was cloned to the pET28b vector after PCR, and the recombinant expression plasmid pET28b-P30 was constructed. Then the recombinants were transformed into E.coli BL21(DE3) after identified by the restriction enzyme digestion, PCR and DNA sequence determination annlysis. A single colony of E.coli BL21(DE3) containing the recombinant plasmid, pET28b-P30 was inoculated in LB culture, then diluted 1∶100 into 2 ml LB culture and induced by 0.2 mmol/L IPTG, and the expression product was identified by SDS-PAGE and Western blot. Results The recombinant plasmid of pET28b-P30 was constructed. ② Plasmid pET28b-P30 could express a specific 30 kDa fusion protein in E.coli BL21(DE3). Conclusions The expression plasmid which contains the gene fragment encoding P30 surface antigen of Toxoplasma gondii has been successfully constructed and is highly expressed in E.coli BL21(DE3) as an inclusion body.