|
|
| |
|
|
码上扫一扫！ |
| 西方蜜蜂AmSTPKD3蛋白的分子特征与基因表达模式 |
|
董舒楠,邹培缘,任亚萍,杜丽婷,李坤泽,臧贺,邱剑丰,周彤,傅佳妮,陈大福,郭睿
|
|
1.福建农林大学 蜂学与生物医药学院，福建 福州 350002;2.天然生物毒素国家地方联合工程试验室，福建 福州 350002;3.福建省蜂疗研究所，福建 福州 350002
|
|
| 摘要: |
| 【目的】对西方蜜蜂（Apis mellifera）的丝氨酸/苏氨酸蛋白激酶（AmSTPKD3）进行生物信息学分析， 测定AmSTPKD3基因在西方蜜蜂工蜂的组织和发育阶段的表达模式，为AmSTPKD3的功能研究提供科学依据。【方法】利用相关生物信息学软件，预测AmSTPKD3的理化性质和分子特征，鉴定西方蜜蜂和其他物种（黑大蜜蜂（Apis laboriosa）、欧洲雄蜂（Bombus terrestris）、大蜜蜂（Apis dorsata）、小蜜蜂（Apis florea）、田野熊蜂（Bombus pascuorum）、东方蜜蜂（Apis cerana）、美洲东部熊蜂（Bombus impatiens）和巴西无刺蜜蜂（Frieseomelitta varia））STPKD3的结构域和保守基序。通过Mega 11.0软件对西方蜜蜂和其他物种的STPKD3进行系统进化分析。采用RT-qPCR方法，检测AmSTPKD3在西方蜜蜂不同组织（触角、毒腺、脑、中肠、脂肪体、表皮和咽下腺）和发育阶段（卵、3日龄幼虫、7和8日龄预蛹、12日龄蛹及1,2,6，12，15和18日龄成虫）的相对表达量。【结果】AmSTPKD3包含3 269个核苷酸，可编码831个氨基酸。AmSTPKD3的分子式为C4141H6511N1155O1283S40，分子质量约为94.29 ku，脂溶系数为77.36，等电点为6.09，平均亲水系数为-0.510，有99个磷酸化位点，但不含跨膜结构域和信号肽。在西方蜜蜂和其他8种蜜蜂的STPKD3中均鉴定到4个相同结构域和5个相同保守基序。西方蜜蜂与东方蜜蜂的STPKD3在进化树上聚为一支。AmSTPKD3在西方蜜蜂工蜂的触角、毒腺、脑、中肠、脂肪体、表皮和咽下腺中差异表达，在咽下腺中的表达量最高，且显著高于脑、中肠、脂肪体和表皮；AmSTPKD3在卵、3日龄幼虫、7和8日龄预蛹及12日龄蛹中差异表达，在卵和8日龄预蛹中的表达量相近，且显著高于3日龄幼虫和12日龄蛹；AmSTPKD3在1,2,6和12日龄成虫体内差异表达，在1日龄成虫体内的表达量最高且显著高于15和18日龄成虫。【结论】AmSTPKD3是潜在的亲水性蛋白和胞内蛋白，西方蜜蜂和其他9种蜂的STPKD3具有较高的保守性，西方蜜蜂与东方蜜蜂STPKD3的同源性最高，AmSTPKD3在西方蜜蜂工蜂的不同组织和发育阶段呈现动态差异表达。 |
| 关键词: 西方蜜蜂 丝氨酸/苏氨酸蛋白激酶 理化性质 分子特征 系统进化 时空表达谱 |
| DOI：DOI:10.13207/j.cnki.jnwafu.2025.06.004 |
| 分类号: |
| 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）；福建农林大学硕士生导师团队项目；福建农林大学科技创新专项基金项目(KFb22060XA)；福建省大学生创新创业训练计划项目（X202310389080,X202310389086） |
|
| Molecular property and gene expression profile of Apis mellifera STPKD3 protein |
|
DONG Shunan1, ZOU Peiyuan1, REN Yaping1, DU Liting1, LI Kunze1, ZANG He1,2,3, QIU Jianfeng1,2,3, ZHOU Tong1, FU Jiani1, CHEN Dafu1,2,4, GUO Rui1,2,4
|
|
1.College of Bee Science and Biomedicine,Fujian Agriculture and Forestry University,Fuzhou，Fujian 350002,China;2.National & Local United Engineering Laboratory of Natural Biotoxin,Fuzhou，Fujian 350002,China;3.Apitherapy Research Institute of Fujian Province,Fu;4.Apitherapy Research Institute of Fujian Province,Fuzhou，Fujian 350002,China
|
| Abstract: |
| 【Objective】In this study,bioinformatic analysis of the Apis mellifera serine/threonine-protein kinase (AmSTPKD3) was performed,followed by determination of the expression profile of AmSTPKD3 in different tissues and developmental stages of Apis mellifera workers,aiming to offer scientific basis for functional investigation of AmSTPKD3.【Method】Relative bioinformatics software were employed to predict the physicochemical characteristics and molecular feature of AmSTPKD3 and to identify structural domains and conserved motifs in STPKD3 from Apis mellifera and other species including Apis laboriosa,Bombus terrestris,Apis dorsata,Apis florea,Bombus pascuorum,Apis cerana,Bombus impatiens and Frieseomelitta varia.Phyletic evolution analysis of STPKD3 in Apis mellifera and other species was performed using Mega 11.0 software.The relative expression level of AmSTPKD3 in Apis mellifera’s various tissues（antennae,venom gland,brain,midgut,fat body,epidermis,and hypopharyngeal gland and developmental stages（egg,3-day-old larva,7- and 8-day-old pre-pupae,12-day-old pupa,and 1-,2-,6-,12-，15-,and 18-day-old worker adults）was evaluated by RT-qPCR.【Result】AmSTPKD3 contained 3 269 nucleotides,encoding a total of 831 amino acids.The molecular formula of AmSTPKD3 was C4141H6511N1155O1283S40,and the molecular weight,lipid solubility coefficient,isoelectric point,average hydrophilicity coefficient were 94.29 ku,77.36,6.09 and -0.510,respectively.AmSTPKD3 included 99 phosphorylation sites,however,no transmembrane domain and signal peptide were detected.The same 4 structural domains and 5 conserved motifs were discovered in STPKD3 from Apis mellifera and 8 other species.The STPKD3 of the Apis mellifera and Apis cerana were grouped into one clade on the phylogenetic tree.AmSTPKD3 was differentially expressed in Apis mellifera worker’s antennae,venom gland,brain,midgut,fat body,epidermis,and hypopharyngeal gland,with the highest expression in hypopharyngeal gland and significantly higher than that in brain,midgut,fat body and epidermis.Additionally,AmSTPKD3 was differentially expressed in egg,3-day-old larva,7- and 8-day-old pre-pupae as well as 12-day-old pupa,and its expression level in egg was similar to that in 8-day-old larva and significantly higher than that in 3-day-old larva and 12-day-old pupa.Moreover,AmSTPKD3 was differentially expressed in 1-,2-,6-,12-day-old adults,and its expression level in 1-day-old adult was the highest and significantly higher than that in 15- and 18-day-old adults.【Conclusion】AmSTPKD3 was a putative hydrophilic and intracellular protein,STPKD3 among Apis mellifera and 9 other bees were highly conservative, and the homology of STPKD3 from Apis mellifera and Apis cerana was the highest.AmSTPKD3 was dynamically and differentially expressed in Apis mellifera workers’ different tissues and developmental stages. |
| Key words: Apis mellifera serine/threonine-protein kinase physicochemical property molecular characteristics phyletic evolution spatio-temporal expression pattern |
|
|
|
|
