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| 西方蜜蜂AKHR基因的生物信息学及表达模式分析 |
| Analysis of bioinformatic and expression pattern of Apis mellifera AKHR gene |
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| DOI： |
| 中文关键词: 西方蜜蜂 脂动激素受体 分子特性 系统进化 表达特征 |
| 英文关键词:Apis mellifera adipokinetic hormone receptor molecular characteristics phyletic evolution expression pattern |
| 基金项目:国家自然科学基金面上项目（32372943，32172792）；国家现代农业产业技术体系建设专项资金项目（CARS-44-KXJ7）；福建省自然科学基金面上项目（2022J01131334）；福建农业职业技术学院科技研究项目（2023JS011）；福建农林大学硕士生导师团队项目（郭睿）；福建农林大学科技创新专项基金（KFb22060XA） |
| Author Name | Affiliation | | LIU Cai-Zhen, FAN Xiao-Xue, WU Bo-Wen, Wu Yang, FU Zhi-Ying, CHEN Da-Fu, GUO Rui | 1. College of Modern Agricultural Engineering, Fuzhou 350000, China
2. College of Bee Science and Biomedicine, Fujian Agriculture and Forestry University, Fuzhou 350002, China
3. National & Local United Engineering Laboratory of Natural Biotoxin, Fuzhou 350002, China
4. Apitherapy Research Institute of Fujian Province, Fuzhou 350002
China |
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| 中文摘要: |
| 本研究对西方蜜蜂Apis mellifera脂动激素受体（Adipokinetic hormone receptor, AKHR）基因AmAKHR CDS区进行RT-PCR扩增和生物信息学分析，并测定了AmAKHR在工蜂不同组织和不同发育阶段的表达模式，旨在为进一步的功能研究提供参考和基础。通过PCR扩增AmAKHR的CDS，使用相关软件预测AmAKHR蛋白的理化性质、分子特征及蛋白互作网络。采用RT-qPCR技术检测AmAKHR在工蜂不同组织和不同发育阶段的相对表达量。结果表明，AmAKHR基因CDS长度为1 050 bp，编码349个氨基酸；AmAKHR的相对分子量约为40.63 kDa，分子式为C1873H2939N471O484S26，含39个磷酸化位点，7个α-跨膜螺旋结构及8个保守基序，不含信号肽，主要分布于质膜；AmAKHR与小蜜蜂Apis florea的AKHR在进化树上聚为一支；AmAKHR的二级结构包含127个无规则卷曲，124个α-螺旋，87条延伸链和11个β-转角；AmAKHR与卵黄原蛋白和神经肽Corazonin等10个蛋白构成1个互作网络；AmAKHR在西方蜜蜂工蜂的毒腺、中肠、咽下腺、脂肪体、脑和触角等6个组织中差异表达，在脂肪体中的表达量最高，而在中肠中的表达量最低；AmAKHR在西方蜜蜂工蜂1日龄幼虫、3日龄幼虫、5日龄幼虫、2日龄预蛹、1日龄蛹中均有表达，且表达量随着日龄的增长而持续上调。研究结果揭示AmAKHR为疏水性蛋白和膜蛋白，并与卵黄原蛋白和神经肽等10个蛋白潜在互作，AmAKHR可能在西方蜜蜂工蜂的能量代谢、脂类分解和变态发育中发挥作用。 |
| 英文摘要: |
| In this study, molecular cloning and bioinformatic analysis of the Apis mellifera gene encoding adipokinetic hormone receptor (AKHR) were conducted, followed by determination of the expression pattern of AmAKHR in various tissues and developmental stages of worker, aiming to offer reference and foundation for further functional investigation. The CDS of AmAKHR was amplified by RT-PCR. Related software were used to predict physicochemical property, molecular characteristics and protein interaction network of AmAKHR. RT-qPCR technology was employed to detect the relative expression level of AmAKHR in different tissues and developmental stages. The results demonstrated that the length of CDS of AmAKHR was about 1 050 bp, encoding 349 amino acids. The molecular weight of AmAKHR was approximately 40.63 kDa, and the molecular formula was C1873H2939N471O484S26. AmAKHR included 39 phosphorylation modification sites, 7 α-transmembrane helical structures and 8 conserved motifs, but there was no signal peptide. AmAKHR was mainly distributed in the plasma membrane. The AmAKHR and the Apis florea AKHR was clustered into one clade on the evolutionary tree. The secondary structure of AmAKHR contained 127 random coils, 124 alpha helices, 87 extended strands and 11 beta turns. AmAKHR formed a interaction network with 10 other proteins such as Vg and Corazonin. AmAKHR was differentially expressed in six tissues of Apis mellifera worker, including venom gland, midgut, hypopharyngenal gland, fat body, brain and antrnna. The expression level of AmAKHR was highest in fat body and lowest in midgut. AmAKHR was expressed in egg, 1-day-old larva, 3-day-old larva, 5-day-old larva, 2-day-old prepupa and 1-day-old pupa, and the expression level was continuously up-regulated as day-old increased. The results reveal that AmAKHR is a kind of hydrophobic protein and membrane protein, AmAKHR potentially interact with 10 proteins such as Vg and Corazonin, AmAKHR is likely to play a role in energy metabolism, lipolysis and metamorphosis of Apis mellifera worker. |
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