关键词: 日本血吸虫, 精氨酸甲基转移酶1, 克隆, 免疫保护, 小鼠

Abstract:

【Objective】 The present study was intend to clone a full length cDNA encoding protein arginine methyltransferase 1 in Schistosoma japonicum, and further access its immunoprotection in mice for schistosomiasis. 【Method】 A full-length cDNA clone encoding a protein arginine methyltransferase was obtained from schistosomula cDNA enriched library, named S.jPRMT1 based on bioinformatics analysis. The expression profiles of SjPRMT1 was determined by real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The open reading frame was then subcloned into a pET28a(+) vector and transformed into competent E.coil/BL21. The recombinant protein was purified and then the immune characters of antigen were accessed by Western blotting. 【Result】 Bioinformatics analysis indicated that the ORF of S.jPRMT1 is 660 base pairs, which is encoding 219 amino acids. Real-time PCR analysis revealed that there is a highest expression in 18 days schistosomula, and slightly lower in 13 days and 23 days parasites, suggesting that S.jPRMT1 has highly expression in schistosomula stage. The recombinant protein showed a good ability to induce antibodies in mice as examined by ELISA. Animal experiment indicated that the administration of recombinant PRMT1 protein in mice resulted in worm burden reduced by 35.07% and egg burden reduced by 48.66%. 【Conclusion】 A full-length cDNA differently expression in schistosomula was obtained. Results demonstrated that the recombinant PRMT1 protein could induce partial protection against schistosomiasis in mice.

Key words: Schistosoma Japonicum, protein arginine methyltransferase PRMT1, subclone, immunoprotection, mice