Cloning, Expression and Antigenicity Analysis of Thioredoxin Peroxidase Gene of Taenia Multiceps
|School||Gansu Agricultural University|
|Course||Preventive Veterinary Medicine|
|Keywords||Long Taenia Brain Coenurus . Thioredoxin peroxidase Recombinant proteins Immunogenicity|
Brain Coenurus disease (Cerebral coenurosis), commonly known as cerebral hydatid disease by bulls Taenia (Taenia multiceps, Tm), larvae - Brain Coenurus the (Coenurus cerebralis) parasites on the host central nervous tissue caused a serious fatal zoonotic parasitic diseases. In China the disease was nationwide distribution, especially in the northwest, north, northeast, vast pastoral areas common, the upward trend in recent years. The disease not only caused huge economic losses to livestock production, and the threat to human health. Thioredoxin peroxidase (Thioredoxin peroxidase, TPx) peroxide thioredoxin (Peroxiredoxin, Prx) of one of the family, is a mercapto Kitt heterosexual antioxidant protein, is used to clear the normal cells of the body generated an excess of reactive oxygen species. The parasite TPx important function of protecting the parasite resistance to the destruction of reactive oxygen species generated from the host body's own metabolism and help the parasite to evade host immune mechanisms. The study according to the pig Taenia TPx gene sequence primers designed from Gansu Jingtai sheep source Coenurus to the clone to TmTPx gene fragments and sequence analysis of the consistency of Taenia showed that in GenBank TaHc2-D11 mRNA sequence (EF420372) 99% the Taenia solium Prx mRNA sequence (AY728092) consistency of 96%. The recombinant expression plasmid pGEX-4T-TmTPx, transformed into E. coli BL21, isopropyl yl-β-D-thiogalactoside (IPTG)-induced expression of recombinant proteins. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-Page) analysis show that the successful expression of a fusion protein of approximately 43 kDa in size. Expression products were purified using the immunizing rabbits, collecting serum antibody responses were measured by ELISA, and the results of the recombinant protein with immune rabbit serum response, antibody levels increased with the increase in the number of immune and rabbit anti TmTPx recombinant protein antibody with the bulls granulosus protoscoleces antigen specifically react better immunogenicity of the recombinant protein. In addition, according to the Taenia TaHc2-D11 mRNA primers were designed using the RT-PCR technique, further amplified to the size of 614 bp TmTPx gene full-length cDNA containing 591 bp open reading frame (ORF) sequence encoding 196 amino acids, the molecular weight of 21.69 kDa, isoelectric point of 7.61. Bioinformatics analysis showed that TmTPx has a typical 2-Cys Prx conserved functional domains in the evolutionary analysis on the tapeworms the known TPx the molecule, found the bulls with tapeworms with Taenia kinship recently, Taenia solium and The fat head tapeworm followed furthest with Echinococcus multi of genetic Echinococcusmultilocularis relationship, which laid the foundation for further study of the function of TmTPx.