猪带绦虫硫氧还蛋白过氧化物酶的原核表达及其生物学特性分析

doi:10.11843/j.issn.0366-6964.2014.09.018

Abstract

Abstract:

Taenia solium (Ts) is an sort of important zoonotic parasite，and thioredoxin peroxidase (TPx) is a protein with important function both in Taenia solium and its larvae Cysticercus cellulosae.The aim of this study was to clarify the biological roles of TsTPx and screen an efficient immunologic target against Cysticercosis.The gene tstpx was obtained by RT-PCR with the total RNA of Taenia solium as template and was confirmed by sequencing.Then the recombinant plasmid pET-30a(+)-tstpx was constructed，subsequently transformed into E.coli BL21，and induced for expression with IPTG.After purification of TsTPx by Ni²⁺-NTA affinity chromatography，the biological properties of TsTPx including antioxidative activity and immunogenicity were analyzed.Protein TsTPx prokaryotically expressed in this study predominantly located in the supernatant as a soluble protein confirmed by SDS-PAGE.Its antioxidative property was proved by，positive correlation of TsTPx concentration to protection percentage of plasmid’s super-coiled DNA structure in metal catalytic oxidation system，where 20 μg•mL^－1 TsTPx achieved 100% protection confirmed by disappearance of nicking slower band in agarose gelelectrophoresis；and also positive correlation of TsTPx concentration to H₂O₂clearance rate in H₂O₂reduction system，where 100 μg•mL^－1 TsTPx achieved 69.9% and 54.5% of H₂O₂clearance with and without dithiothreitol，respectively.Its antigenicity was demonstrated by special recognition between Cysticercus positive serum from pig in Western blotting and antibody titers up to 1：320 000 detected with indirect ELISA after TsTPx vaccine was prepared and injected into rabbits.The results showed that the prokaryotically expressed TsTPx in present study was biotically active and showing satisfied antigenicity，which laid the foundation for a comprehensive study of its biological function and for the development of a vaccine against cysticercosis.

CLC Number:

S852.734

YIN Cai，LUO Xue-nong，WANG Shuai，CHEN Guang-li，ZENG Qiao-ying，CAI Xue-peng. Prokaryotic Expression and Biological Properties of Thioredoxin Peroxidase from Taenia solium[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(9): 1512-1517.

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Prokaryotic Expression and Biological Properties of Thioredoxin Peroxidase from Taenia solium

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