中华蜜蜂PP2A基因的时空表达谱及蛋白分子特征
Spatio-temporal expression profile and molecular properties of the Apis cerana cerana PP2A gene
康 婧1** 陈 颖1** 孙恺玥1 王勇杰1 臧 贺1, 2, 3 冯睿蓉4 付中民1 陈大福1, 2, 3 邱剑丰1, 2, 3*** 郭 睿1, 2, 3***
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DOI:10.7679/j.issn.2095-1353.2025.086
作者单位:1. 福建农林大学蜂学与生物医药学院,福州 350002;2. 天然生物毒素国家地方联合工程实验室,福州 350002; 3. 福建农林大学蜂疗研究所,福州 350002;4. 福建农业职业技术学院,福州 350000
中文关键词:中华蜜蜂;蛋白磷酸酶2A;时空表达谱;蛋白分子特征
英文关键词:Apis cerana cerana; protein phosphatase 2A; spatio-temporal expression profile; molecular features of proteins
中文摘要:【目的】 本研究旨在通过测定中华蜜蜂Apis cerana cerana蛋白磷酸酶2A(Protein phosphatase 2A , PP2A)基因AcPP2A的时空表达谱和解析AcPP2A蛋白的理化性质和分子特征,为进一步研究PP2A在中华蜜蜂生长发育中的功能提供参考和基础。【方法】 通过RT-qPCR检测AcPP2A在中华蜜蜂工蜂的触角、中肠、脑、毒腺、表皮、咽下腺和脂肪体等组织,卵、幼虫、预蛹和蛹及不同日龄工蜂成虫中的相对表达量。使用NCBI网站上的ORF工具,Expasy网站上的Protparam、ProtScale和SWISS-model软件,SignalP 4.1 Server、NetPhos 3.1 Server、TMHMM和SOPMA等软件预测AcPP2A蛋白的分子特征和理化性质,鉴定中华蜜蜂和其他物种PP2A的保守基序和结构域,并构建系统进化树。【结果】 AcPP2A含有5 174个核苷酸,可编码475个氨基酸;AcPP2A的蛋白分子量约为54.78 kD,脂溶系数为78.80,等电点为5.97,分子式为C2411H3781N671O742S23,平均亲水系数为﹣0.521;AcPP2A不含跨膜结构域和信号肽。中华蜜蜂、的PP2A含有相同的6个保守基序(Motif1、Motif2、Motif3、Motif4、Motif5和Motif6)和1个相同的结构域(CDC55超家族)。中华蜜蜂与橡子蚂蚁的PP2A在进化树上聚为一支。【结论】 AcPP2A是潜在的酸性蛋白、亲水性蛋白和非跨膜胞内蛋白,具有较高的保守性;AcPP2A的表达存在组织及发育阶段的广泛性和差异性,在毒腺和卵发育中可能发挥重要功能。
英文摘要:
[Aim] To determine the spatio-temporal
expression profile of the Apis cerana cerana protein phosphatase 2A (PP2A) gene, analyze its molecular
characteristics and physical and chemical properties in order to provide a
reference for further research. [Methods] The relative expression of AcPP2A in different tissues, including the antennae, midgut, brain, venom gland,
cuticle, hypopharyngeal gland, and fat body, were determined using RT-qPCR, as were its relative expression in eggs,
larvae, pre-pupae, pupae and adult workers of different ages. Using the ORF
tool on the NCBI website, Protparam, ProtScale, and SWISS-model software on the
Expasy website, SignalP 4.1 Server, NetPhos 3.1 Server, TMHMM, and SOPMA, and
other software related software Predicting molecular features of the AcPP2A
protein and the physicochemical properties, identify conserved motifs and
structural domains of PP2A in A. c. cerana and other species, and
construct a phylogenetic tree. [Results] AcPP2A contains 5 174
nucleotides, encodes 475 amino acids and has a protein molecular weight of
about 54.78 kD. It has a lipid solubility coefficient of 78.80, an isoelectric
point of 5.97, a molecular formula of C2411H3781N671O742S23,
and an average hydrophilic coefficient of ﹣0.521. AcPP2A
does not have a transmembrane domain or signaling peptide. PP2A of A. c.
cerana, Melipona quadrifasciata, Apis mellifera, Bombus
impatiens, Temnothorax curvispinosus, Megachile rotundata, B.
pyrosoma, Frieseomelitta varia, Acromyrmex echinatior, Habropoda
laboriosa and Lingula anatina, all have the same six conserved
motifs (Motif1, Motif2, Motif3, Motif4, Motif5 and Motif6) and one identical
domain (the CDC55 superfamily). PP2A of A. c. cerana and T.
curvispinosus cluster on the same branch of the phylogenetic tree. [Conclusion] AcPP2A appears to be an acidic, hydrophilic,
non-transmembrane, highly conserved, intracellular protein. Its expression
varies widely across different tissues and developmental stages, but it may
play an important role in the development of the venom gland and eggs.