Identification And Characterization Of Novel Peroxiredoxin Prx And PrxQ Of Babesia Microti

In recent years, Babesiosis microti has become a new type of human pathogens, and in many parts of the world have been reported sporadic cases.Parasite peroxiredoxins(Prxs) is a class of protein which expressed rich in cells. Prxs antioxidant protein family like other proteases, such as SOD, Gpx, Trx and Trx R all play important role in protecting vivo biological macromolecules from the aspect of negative factors. Prxs reduction of hydrogen peroxide is mainly dependent on its highly conserved redox activity Cys. Prxs is divided into Prx1, Prx5, Prx6, TPX, Prx Q and Ahp E six subfamilies. In this reaserach, we were focus on Prx1 and Prx Q subfamily. Although the Prxs genome of this parasite has been sequenced, but their biological properties are still incomplete.In this study, we used molecular biology techniques and obtained total length of gene sequence of Prx1 and Prx Q. The total length of gene sequence of Prx1 is 594 bp, encoding 194 amino acids, having a open reading frame of ORF and using software speculate Prx1 protein size is approximately 23 k Da. Prx Q total gene sequences of 537 bp in length, having an ORF encoding 194 amino acids of the open reading frame, which has a conserved sequence in the signal peptide sequence, using software presumed Prx Q protein size of about 21 k Da. And successfully constructed with p ET-30 a as a carrier of recombinant plasmid successfully transformed into BL21 competent cells of prokaryotic expression system. By inducing we get a lot of recombinant protein and in line with further experiments.Prx1 and Prx Q activity analysis showed that the protein have proven Prx1 and Prx Q have activity but Prx1 more active. In the recombinant plasmid we found, Prx Q sequence has a signal peptide, which may determine different distribution location of Prx1 and Prx Q in the cell, so that the two proteins play different functions. Furthermore, proof has been successfully prepared Prx1 and Prx Q antiserum identified by Western blot and indirect immunofluorescence test(IFAT) analysis, and Western blot analysis proved that the parasite Prx1 and Prx Q natural protein in monomeric form of the parasite to exert antioxidation. Prx1 and Prx Q abundantly expressed when the parasite in red blood cells split peak. IFAT proof Prx1 and Prx Q abound with natural proteins in the cytoplasm. In this paper, we were focuse on the function and biological significance of Babesia microti peroxiredoxins, and explore related biological characteristics of Prx of Babesia microti. Desirable to provide a reference for the study of pathogenesis of B. microti and the development of protozoosis vaccine, and provide the basis for further research.