Expression of the P30 gene of Toxoplasma gondii as MBP fusion protein.

Aim Construction of Prokaryotic expression vector contained a gene encoding P30 gene of T gondii and expression in E coli Methods According to the published P30 sequence,a pair of primer was designed and at the 5’ end E coRI and SalI enzyme sites were added respectively Using PCR technique,a fragment of P30 gene was obstained by amplification of genomic DNA of tachyzoites:By direct cloning,P30 gene was inserted into pMALP 2 plasmid,then transformed into E coli DH5 a The recombinant clones were induced by IPTG concentration A high level expression of MBP fusion protein was analyzed by means of SDS-PAGE and Western-blot Results The gene encoding P30 was successfully amplified from the genomic DNA of RH strain of Toxoplasma gondii by PCR,and the recombinant plasmid pMALP 2-P30was constructed successfully The results of SDS-PAGE and Western-blot revealed that the molecular weight of recombinant protein MBP/P30 was approximately 77 5KD,and P30 is 34 5KD out of MBP 43KD and can be specifically recognized by rat antiserum of Toxoplasma Conclusion P30 gene was obtained from genomic DNA of Toxoplasma and pMALP 2-P30 recombinant was successfully constructed The expression of P30 fusion protein will be used to purify P30 protein and study it's immunity in to animal