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,SignalP4.1 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、无刺蜂Melipona quadrifasciata、西方蜜蜂Apis mellifera、美洲东北熊蜂Bombus impatiens、橡子蚂蚁Temnothorax curvispinosus、苜蓿切叶蜂Megachile rotundata、火红熊蜂Bombus pyrosoma、巴西无刺蜂Frieseomelitta varia、顶切叶蚁Acromyrmex echinatior、东南蓝莓蜂Habropoda laboriosa、鸭嘴海豆芽Lingula anatina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。
[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 5174 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.
作者
康婧
陈颖
孙恺玥
王勇杰
臧贺
冯睿蓉
付中民
陈大福
邱剑丰
郭睿
KANG Jing;CHEN Ying;SUN Kai-Yue;WANG Yong-Jie;ZANG He;FENG Rui-Rong;FU Zhong-Min;CHEN Da-Fu;QIU Jian-Feng;GUO Rui(College of Bee Science and Biomedicine,Fujian Agriculture and Forestry University,Fuzhou 350002,China;National&Local United Engineering Laboratory of Natural Biotoxin,Fuzhou 350002,China;Apitherapy Research Institute of Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Vocational College of Agriculture,Fuzhou 350000,China)
出处
《应用昆虫学报》
2025年第4期915-924,共10页
Chinese Journal of Applied Entomology
基金
国家自然科学基金面上项目(32172792,32372943)
国家现代农业产业技术体系建设专项资金(CARS-44-KXJ7)
福建省自然科学基金面上项目(2023J01133)
福建省教育厅中青年教师教育科研项目(JAT231226)
福建农林大学硕士生导师团队项目(郭睿)。
关键词
中华蜜蜂
蛋白磷酸酶2A
时空表达谱
蛋白分子特征
Apis cerana cerana
protein phosphatase 2A
spatio-temporal expression profile
molecular features of proteins
作者简介
共同第一作者:康婧,E-mail:kj000213@163.com;共同第一作者:陈颖,E-mail:13979562297@163.com;共同通讯作者:邱剑丰,E-mail:jfqiu@fafu.edu.cn;共同通讯作者:郭睿,E-mail:ruiguo@fafu.edu.cn。
