The molecular characterization and immune protection of microneme 2 of Eimeria acervulina

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• 英文论文题名：The molecular characterization and immune protection of microneme 2 of Eimeria acervulina
• 论文作者：Zhenchao Zhang ; Lianrui Liu ; Jingwei Huang ; Ruofeng Yan ; Lixin Xu ; Xiaokai Song ; Xiangrui Li
• 英文论文作者：Zhenchao Zhang ; Lianrui Liu ; Jingwei Huang ; Ruofeng Yan ; Lixin Xu ; Xiaokai Song ; Xiangrui Li ; College of Veterinary Medicine ; Nanjing Agricultural University
• 年：2016
• 作者机构：College of Veterinary Medicine, Nanjing Agricultural University;
• 英文论文关键词：E ; acervulina ; EaMIC2 ; Immunogenicity
• 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：S852.723
• 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：152k
• 原文格式：D
• 会议级别：全国

摘要

Eimeria acervulina(E. acervulina) is an intestinal protozoon and few of its antigen genes were researched. In this study, the gene of E. acervulina microneme protein 2(Ea MIC2) was cloned and characterized. The degenerate primers for the conserved sequence of microneme protein 2 were designed based on Eimeria tenella microneme 2(EtMIC2)(AF111839.1), Eimeria maxima microneme 2(Em MIC2)(FR718971.1) and E. brunetti microneme 2(Eb MIC2)(AB723700.1) to amplify the conserved sequence of microneme protein 2. According to the conserved sequence, specific primers for the rapid amplification of cD NA ends(RACE) were designed to amplify the 3'- and 5'-ends of Ea MIC2. The full length c DNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. The sequence analysis revealed that the opening reading frame(ORF) of Ea MIC2 was 882 bp and encoded a protein of 293 amino acids with 29.81 k Da. The most part of the mature protein(base: 82-882 bp, amino acid: 28-293aa) of Ea MIC2(mpm Ea MIC2) was inserted into p Cold TF to produce recombinant mpm Ea MIC2. Western blotting assay was used to analysis the immunogenicity of mpm Ea MIC2 and the result showed that the recombinant protein was successfully recognized by the sera of chickens experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of mpmE a MIC2. Immunofluorescence analysis using antibody against recombinant protein mpmE a MIC2 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of mpm Ea MIC2 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens. All the above results suggested that the Ea MIC2 was a novel E. acervulina antigen and could be an effective candidate for the development of new vaccine against this parasite.
Eimeria acervulina(E. acervulina) is an intestinal protozoon and few of its antigen genes were researched. In this study, the gene of E. acervulina microneme protein 2(Ea MIC2) was cloned and characterized. The degenerate primers for the conserved sequence of microneme protein 2 were designed based on Eimeria tenella microneme 2(EtMIC2)(AF111839.1), Eimeria maxima microneme 2(Em MIC2)(FR718971.1) and E. brunetti microneme 2(Eb MIC2)(AB723700.1) to amplify the conserved sequence of microneme protein 2. According to the conserved sequence, specific primers for the rapid amplification of cD NA ends(RACE) were designed to amplify the 3'- and 5'-ends of Ea MIC2. The full length c DNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. The sequence analysis revealed that the opening reading frame(ORF) of Ea MIC2 was 882 bp and encoded a protein of 293 amino acids with 29.81 k Da. The most part of the mature protein(base: 82-882 bp, amino acid: 28-293aa) of Ea MIC2(mpm Ea MIC2) was inserted into p Cold TF to produce recombinant mpm Ea MIC2. Western blotting assay was used to analysis the immunogenicity of mpm Ea MIC2 and the result showed that the recombinant protein was successfully recognized by the sera of chickens experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of mpmE a MIC2. Immunofluorescence analysis using antibody against recombinant protein mpmE a MIC2 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of mpm Ea MIC2 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens. All the above results suggested that the Ea MIC2 was a novel E. acervulina antigen and could be an effective candidate for the development of new vaccine against this parasite.

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