Identification of a thioredoxin reductase(TrxR) from Babesia microti
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- 英文论文题名：Identification of a thioredoxin reductase(TrxR) from Babesia microti
- 论文作者：Haiyan Gong ; Shaoruo Zhao ; Kang Xiong ; Hao Zhang ; Houshuang Zhang ; Yongzhi Zhou ; Jie Cao ; Jinlin Zhou
- 英文论文作者：Haiyan Gong ; Shaoruo Zhao ; Kang Xiong ; Hao Zhang ; Houshuang Zhang ; Yongzhi Zhou ; Jie Cao ; Jinlin Zhou ; Key Laboratory of Animal Parasitology of Ministry of Agriculture ; Shanghai Veterinary Research Institute ; Chinese Academy of Agricultural Sciences
- 年：2016
- 作者机构：Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences;
- 英文论文关键词：Babesia microti ; transcriptome ; Trx R ; reductive activity
- 会议召开时间：2016-08-08
- 会议录名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会论文摘要集
- 英文会议录名称：The Abstracts of the Tenth Symposium for Yong Scientists of China Society of Parasitology
- 语种：英文
- 分类号：S852.7
- 学会代码：JISC
- 会议名称：中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会
- 会议地点：中国四川成都
- 主办单位：中国动物学会寄生虫学专业青年委员会
- 学会名称：中国动物学会寄生虫学专业委员会
- 页数：1
- 文件大小：154k
- 原文格式：D
- 会议级别：全国
摘要
Babesia, a kind of tick-borne pathogen to infect mammals including human, causes a great lose for the husbandry and threats the health of human. The drugs for babesiosis always bring side effect and drug resistance, thus the development of new drugs or vaccines seems important and necessary. As it lives in the red blood cell and surrounded by reactive oxygen species(ROS), Babesia involved the thioredoxin(Trx) system to deal with the oxidative stress. Aims: In order to explore the Trx system from Babesia, the transcriptome of the parasite was sequenced, and from which a Trx reductase(TrxR) was isolated and characterized. Method: The full length of TrxR was obtained by 3' and 5' RACE. The gene was ligated into pE T-30 a and the recombinant protein was purified for antibody preparation and enzymetic activity analysis. The native TrxR from B. microti(Bmi TrxR) was identified by Western blotting and IFAT. Results: TrxR is a putative 553 amino acids protein with a full length of 1766 bp and a signal peptide of 18 amino acids. The deduced protein sequence showed a highest homology to B. bovis TrxR(XP_001608869, 58%), and 44% to human TrxR. The recombinant protein of TrxR showed reductive activity to DTNB, which is concentration dependent and can be inhibited by Auranofin. The optimum p H value for the reaction is 6.5. The native protein Bmi TrxR with a molecular size of 58.4 k Da was detected with the mouse anti-Bmi TrxR polyclonal serum by western blotting. IFAT result indicates that Bmi TrxR may localize in the cytoplasm of B. microti. Conclusion: A TrxR from B. microti was isolated and functionally identified in the present study. It provided information for the understanding the mechanism of Trx system and for the development of novel drugs to prevent B. microti infection.
Babesia, a kind of tick-borne pathogen to infect mammals including human, causes a great lose for the husbandry and threats the health of human. The drugs for babesiosis always bring side effect and drug resistance, thus the development of new drugs or vaccines seems important and necessary. As it lives in the red blood cell and surrounded by reactive oxygen species(ROS), Babesia involved the thioredoxin(Trx) system to deal with the oxidative stress. Aims: In order to explore the Trx system from Babesia, the transcriptome of the parasite was sequenced, and from which a Trx reductase(TrxR) was isolated and characterized. Method: The full length of TrxR was obtained by 3' and 5' RACE. The gene was ligated into pE T-30 a and the recombinant protein was purified for antibody preparation and enzymetic activity analysis. The native TrxR from B. microti(Bmi TrxR) was identified by Western blotting and IFAT. Results: TrxR is a putative 553 amino acids protein with a full length of 1766 bp and a signal peptide of 18 amino acids. The deduced protein sequence showed a highest homology to B. bovis TrxR(XP_001608869, 58%), and 44% to human TrxR. The recombinant protein of TrxR showed reductive activity to DTNB, which is concentration dependent and can be inhibited by Auranofin. The optimum p H value for the reaction is 6.5. The native protein Bmi TrxR with a molecular size of 58.4 k Da was detected with the mouse anti-Bmi TrxR polyclonal serum by western blotting. IFAT result indicates that Bmi TrxR may localize in the cytoplasm of B. microti. Conclusion: A TrxR from B. microti was isolated and functionally identified in the present study. It provided information for the understanding the mechanism of Trx system and for the development of novel drugs to prevent B. microti infection.
Babesia, a kind of tick-borne pathogen to infect mammals including human, causes a great lose for the husbandry and threats the health of human. The drugs for babesiosis always bring side effect and drug resistance, thus the development of new drugs or vaccines seems important and necessary. As it lives in the red blood cell and surrounded by reactive oxygen species(ROS), Babesia involved the thioredoxin(Trx) system to deal with the oxidative stress. Aims: In order to explore the Trx system from Babesia, the transcriptome of the parasite was sequenced, and from which a Trx reductase(TrxR) was isolated and characterized. Method: The full length of TrxR was obtained by 3' and 5' RACE. The gene was ligated into pE T-30 a and the recombinant protein was purified for antibody preparation and enzymetic activity analysis. The native TrxR from B. microti(Bmi TrxR) was identified by Western blotting and IFAT. Results: TrxR is a putative 553 amino acids protein with a full length of 1766 bp and a signal peptide of 18 amino acids. The deduced protein sequence showed a highest homology to B. bovis TrxR(XP_001608869, 58%), and 44% to human TrxR. The recombinant protein of TrxR showed reductive activity to DTNB, which is concentration dependent and can be inhibited by Auranofin. The optimum p H value for the reaction is 6.5. The native protein Bmi TrxR with a molecular size of 58.4 k Da was detected with the mouse anti-Bmi TrxR polyclonal serum by western blotting. IFAT result indicates that Bmi TrxR may localize in the cytoplasm of B. microti. Conclusion: A TrxR from B. microti was isolated and functionally identified in the present study. It provided information for the understanding the mechanism of Trx system and for the development of novel drugs to prevent B. microti infection.
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