细粒棘球绦虫MAPⅡ生物信息学分析及其在虫体不同阶段表达差异研究

doi:10.16431/j.cnki.1671-7236.2025.07.029

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (7): 3286-3296.doi: 10.16431/j.cnki.1671-7236.2025.07.029

细粒棘球绦虫MAPⅡ生物信息学分析及其在虫体不同阶段表达差异研究

陈旭珂^1,2, 普娜^1,2, 张玉霞^1,2, 赵文卿², 赵家欣^1,2, 黄俊程², 魏琳颖^1,2, 张艳艳², 孙艳², 薄新文², 王正荣²

1. 石河子大学动物科技学院, 石河子 832000;
2. 新疆农垦科学院畜牧兽医研究所, 省部共建绵羊遗传改良与健康养殖国家重点实验室, 石河子 832000

修回日期:2024-11-28 出版日期:2025-07-05 发布日期:2025-07-01
通讯作者: 薄新文, 王正荣 E-mail:851661263@qq.com;wzrtiger@sina.com
作者简介:陈旭珂，E-mail:chenxuke1@outlook.com。
基金资助:
国家自然科学基金(32360887、31860701)；新疆生产建设兵团国际科技合作(2021BC008)；省部共建绵羊遗传改良与健康养殖国家重点实验室重大专项(2021ZD02)；兵团农业科技创新工程专项(NCG202213)

Bioinformatics Analysis of Methionine Aminopeptidase Ⅱ of Echinococcus granulosus and Its Differential Expression in Different Stages of the Worm

CHEN Xuke^1,2, PU Na^1,2, ZHANG Yuxia^1,2, ZHAO Wenqing², ZHAO Jiaxin^1,2, HUANG Juncheng², WEI Linying^1,2, ZHANG Yanyan², SUN Yan², BO Xinwen², WANG Zhengrong²

1. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;
2. State Key Laboratory of Sheep Genetic Improvement and Healthy Breeding, Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural Reclamation Sciences, Shihezi 832000, China

Revised:2024-11-28 Online:2025-07-05 Published:2025-07-01

摘要/Abstract

摘要： 【目的】研究甲硫氨酸氨基肽酶Ⅱ(methionine aminopeptidase Ⅱ，MAPⅡ)的分子特征、反应原性及其在虫体不同发育阶段的表达情况，提供一种对终末宿主具有保护作用的疫苗候选抗原。【方法】对MAPⅡ蛋白进行生物信息学分析，包括系统进化树构建、理化性质分析及信号肽、跨膜区、抗原表位、磷酸化和糖基化位点、二级结构、三级结构预测。通过原核系统表达MAPⅡ蛋白并验证重组蛋白的抗原性，同时进行虫体不同发育阶段转录水平分析。【结果】细粒棘球绦虫MAPⅡ蛋白与终末宿主犬、中间宿主绵羊中 MAPⅡ蛋白亲缘关系较远。MAPⅡ蛋白由453个氨基酸组成，分子质量为51 023.75 u，不稳定系数为36.6，为稳定蛋白；平均疏水性指数为－0.609，亲水性较好。不存在信号肽与跨膜区。MAPⅡ蛋白具有良好的B细胞线性表位，共有38个磷酸化位点，1个N-糖基化位点。MAPⅡ蛋白二级结构以α-螺旋和无规则卷曲为主，占比分别为33.55%和51.88%；三级结构预测显示其为单体蛋白，GMQE值为0.85。通过原核系统表达出的MAPⅡ重组蛋白能与感染细粒棘球绦虫犬的阳性血清结合，抗原性良好。MAPⅡ蛋白在原头蚴阶段和成虫阶段均有表达，且成虫阶段表达量显著高于原头蚴阶段(P＜0.05)。【结论】 MAPⅡ蛋白结构稳定，具有优势抗原表位，MAPⅡ重组蛋白免疫原性良好，其在细粒棘球绦虫不同发育阶段特别是成虫阶段发挥着重要作用，可作为细粒棘球绦虫终末宿主疫苗候选抗原。

关键词: 细粒棘球绦虫; 甲硫氨酸氨基肽酶Ⅱ(MAPⅡ); 生物信息学; 原核表达; 转录水平

Abstract: 【Objective】 The purpose of this experiment was to study the molecular characteristics,reactogenicity and expression of methionine aminopeptidase Ⅱ (MAP Ⅱ) in different stages of Echinococcus granulosus development,and provide a vaccine candidate antigen with protective effect on the definitive host. 【Method】 Bioinformatics analysis of MAP Ⅱ protein was carried out,including phylogenetic tree construction,physical and chemical properties analysis,and prediction of signal peptide,transmembrane region,antigen epitopes,phosphorylation and glycosylation sites,and secondary and tertiary structures.The prokaryotic system was used to express MAP Ⅱ protein and the antigenicity of the recombinant protein was verified.At the same time,the transcript levels of different developmental stages of Echinococcus granulosus were analyzed. 【Result】 The MAP Ⅱ protein of Echinococcus granulosus was distantly related to that of the definitive host dog and the intermediate host sheep.MAPⅡ was composed of 453 amino acids with a molecular weight of 51 023.75 u.The instability coefficient was 36.6,indicating Map Ⅱ was a stable protein.The average hydrophobicity index was －0.609 and the hydrophilicity was good.There was no signal peptide and transmembrane region in MAP Ⅱ protein.Map Ⅱ protein had a good B cell linear epitope,with a total of 38 phosphorylation sites and 1 N-glycosylation site.The secondary structure of MAPⅡ protein was dominated by alpha helix and random coil,accounting for 33.55% and 51.88%,respectively.The tertiary structure prediction showed that it was a monomeric protein,and the GMQE value was 0.85.The MAP Ⅱ recombinant protein expressed by prokaryotic system could bind to the positive serum of dogs infected with Echinococcus granulosus,and had good antigenicity.MAP Ⅱ protein was expressed in both the protoscoleces stage and the adult stage,and the expression in the adult stage was significantly higher than that in the protoscoleces stage (P＜0.05). 【Conclusion 】 Map Ⅱ protein had stable structure and dominant antigen epitopes.The immunogenicity of MAP Ⅱ recombinant protein was good.It played an important role in different developmental stages of Echinococcus granulosus,especially in adult stages,and could be used as a vaccine candidate antigen for definitive host of Echinococcus granulosus.

Key words: Echinococcus granulosus; methionine aminopeptidase Ⅱ(MAPⅡ); bioinformatics; prokaryotic expression; transcription level

中图分类号:

S852.73^＋4

陈旭珂, 普娜, 张玉霞, 赵文卿, 赵家欣, 黄俊程, 魏琳颖, 张艳艳, 孙艳, 薄新文, 王正荣. 细粒棘球绦虫MAPⅡ生物信息学分析及其在虫体不同阶段表达差异研究[J]. 中国畜牧兽医, 2025, 52(7): 3286-3296.

CHEN Xuke, PU Na, ZHANG Yuxia, ZHAO Wenqing, ZHAO Jiaxin, HUANG Juncheng, WEI Linying, ZHANG Yanyan, SUN Yan, BO Xinwen, WANG Zhengrong. Bioinformatics Analysis of Methionine Aminopeptidase Ⅱ of Echinococcus granulosus and Its Differential Expression in Different Stages of the Worm[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3286-3296.

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细粒棘球绦虫MAPⅡ生物信息学分析及其在虫体不同阶段表达差异研究

Bioinformatics Analysis of Methionine Aminopeptidase Ⅱ of Echinococcus granulosus and Its Differential Expression in Different Stages of the Worm

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