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旨在解析东方蜜蜂(Apis cerana)抗细胞凋亡E3泛素蛋白连接酶1(apoptosis-resistant E3 ubiquitin protein ligase 1,Ac AREL1)的理化特性和保守性,并检测Ac AREL1基因应答真菌侵染的表达模式,为其功能研究提供参考。针对Ac AREL1的编码序列(coding sequence, CDS),采用RT-PCR进行分段克隆。利用生物信息学网站和专用软件对Ac AREL1蛋白的理化特性、分子结构、保守基序、结构域进行分析,并探究其系统进化。运用RT-qPCR方法检测东方蜜蜂工蜂受东方蜜蜂微孢子虫(Nosema ceranae)侵染时Ac AREL1的表达模式。成功实现Ac AREL1的编码序列克隆; Ac AREL1含有954个氨基酸,分子质量约为107.55 k Da,平均亲水系数为-0.218,含有典型的信号肽和2个跨膜结构域。Ac AREL1的二级结构含有415个α-螺旋,394个无规卷曲和145条延伸链; Ac AREL1与三级结构模板A0A482X258.1. A的同源性达72.17%。Ac AREL1可能主要定位于内质网。东方蜜蜂、西方蜜蜂(Apis mellifera)、小蜜蜂(Apis florea)、大蜜蜂(Apis dorsata)、无刺蜂(Frieseomelitta varia)、亨特大黄蜂(Bombus huntii)、黄脸熊蜂(Bombus vosnesenskii)、毛眼林蚁(Formica exsecta)和麦茎蜂(Cephus cinctus)的AREL1均含有HECTc和Filamin superfamily结构域及5个相同的保守基序;进化树的分支结构中东方蜜蜂和西方蜜蜂的AREL1归为同一支。东方蜜蜂微孢子虫侵染后,工蜂中肠中Ac AREL1的表达下调; Ac AREL1在1 dpi(1 day post-infection)的表达水平最高,在6 dpi的表达水平最低。本研究首次获得东方蜜蜂AREL1全长序列,分析发现Ac AREL1是潜在的跨膜和亲水性蛋白,跨物种的保守性高;东方蜜蜂微孢子虫侵染后,东方蜜蜂工蜂中肠中Ac AREL1的表达被抑制。研究结果为后续的功能研究提供了基础。
Abstract：The aim of this study was to analyze the physical and chemical characteristics and conservation of apoptotic-resistant E3 ubiquitin protein ligase 1 in Apis cerana(Ac AREL1), and to investigate the expression pattern of the Ac AREL1 gene in response to fungal infection, thereby providing a reference for further functional studies. The coding sequence(CDS) of Ac AREL1 was cloned segmentally using RT-PCR. Bioinformatics tools and specialized software were employed to analyze the physicochemical properties, molecular structure, conserved motifs, and domains of the Ac AREL1 protein, as well as to explore its phylogenetic evolution. RT-qPCR was used to determine the expression pattern of Ac AREL1 in workers of Apis cerana following infection with Nosema ceranae. The CDS of Ac AREL1 was successfully cloned. Ac AREL1 consisted of 954 amino acids, with a molecular weight of approximately 107. 55 k Da and an average hydrophilicity coefficient of-0. 218. It contained a typical signal peptide and two transmembrane domains. The secondary structure of Ac AREL1 comprised 415 α-helices, 394 random coils, and 145 extended strands. Ac AREL1 showed 72. 17% homology with the tertiary structure template A0A482X258. 1. A, and was predicted to be primarily localized in the endoplasmic reticulum.AREL1 proteins from Apis cerana, Apis mellifera, Apis florea, Apis dorsata, Frieseomelitta varia, Bombus huntii, Bombus vosnesenskii,Formica exsecta, and Cephus cinctus all contained HECTc and Filamin superfamily domains, as well as five identical conserved motifs.In the phylogenetic tree, AREL1 from Apis cerana and Apis mellifera clustered within the same branch. After infection with Nosema ceranae, the expression of Ac AREL1 in the midgut of worker bees was down-regulated, with the highest expression level observed at one day post-infection(1 dpi) and the lowest at 6 dpi. This study firstly obtained the full-length sequence of Ac AREL1. Analysis revealed that Ac AREL1 was a potential transmembrane and hydrophilic protein with high conservation across species. The expression of Ac AREL1 in the midgut of Apis cerana workers was suppressed following microsporidian infection. These findings provide a foundation for subsequent functional research.
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基本信息:
DOI：10.13417/j.gab.045.000600
中图分类号:S895
引用信息:
[1]樊念,李想,杨雪,等.东方蜜蜂AREL1基因的克隆、生物信息学及其在东方蜜蜂微孢子虫侵染中的表达模式[J].基因组学与应用生物学,2026,45(03):600-609.DOI:10.13417/j.gab.045.000600.
基金信息:
国家自然科学基金面上项目(32372943,32172792); 国家现代农业蜂产业技术体系专项资金(CARS-44-KXJ7); 福建农林大学科技创新专项基金(KFb22060XA); 福建省大学生创新创业训练计划项目(202510389433,202510389450)共同资助
2026-06-25
2026-06-25