东方蜜蜂微孢子虫RRDRP的分子特性及系统进化分析  

Analysis on molecular characteristics and phyletic evolution of Nosema ceranae RRDRP

作　　者：张凯遥 冯佩林 宓诗雨 郭思佳 张艺琼 荆欣 陈大福[1,2,3] 郭睿 付中民[1,2,3] ZHANG Kaiyao;FENG Peilin;FU Shiyu;GUO Sijia;ZHANG Yiqiong;JING Xin;CHEN Dafu;GUO Rui;FU Zhongmin(College of Bee Science and Biomedicine,Fujian Agriculture and Forestry University;National&Local United Engineering Laboratory of Natural Biotoxin,Fujian Agriculture and Forestry University;Apitherapy Research Institute of Fujian Province,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学蜂学与生物医药学院 [2]福建农林大学天然生物毒素国家地方联合工程实验室 [3]福建省蜂疗研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2024年第3期371-376,共6页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(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项目(2022J01131334、2023J01133656);福建农林大学硕士生导师团队项目(郭睿);福建省大学生创新创业训练计划项目(202310389027、X202310389084)。

摘　　要：【目的】通过对东方蜜蜂微孢子虫和其他物种的Rad18-like重组与DNA修复蛋白(Rad18-like recombination and DNA repair protein,RRDRP)进行系统进化分析,为进一步研究其功能提供理论依据。【方法】利用Protparam、ProtScale和SWISS-model等软件分析RRDRP的理化性质、亲水性和三级结构。利用SignalP 4.1 Server、NetPhos 3.1 Server、TMHMM和SOPMA等软件预测RRDRP的信号肽、磷酸化位点、跨膜结构域及二级结构。使用PSORTⅡ软件预测RRDRP蛋白的亚细胞定位。使用MEME软件预测东方蜜蜂微孢子虫和其他物种RRDRP的保守基序。通过Mega 11.0软件对东方蜜蜂微孢子虫和其他物种的RRDRP进行氨基酸序列多重比对,并采用邻接法构建系统进化树。【结果】东方蜜蜂微孢子虫RRDRP基因含有2928个核苷酸,可编码975个氨基酸,分子质量约11.50 ku,脂溶系数93.58,等电点6.63,分子式C_(5135)H_(8253)N_(1377)O_(1545)S_(34),平均亲水系数为-0.627,可同时定位于细胞核、细胞质、过氧化物酶体和细胞骨架;RRDRP包含55个磷酸化位点、678个α-螺旋、99条延长链、22个β-转角及176个无规则卷曲;RRDRP不含信号肽与跨膜结构域。东方蜜蜂微孢子虫、兔脑炎微孢子虫、颗粒病微粒子虫等11个物种的RRDRP中均含有5个相同的保守基序(Motif 1、Motif 2、Motif 3、Motif 4和Motif 5)。东方蜜蜂微孢子虫与颗粒病微粒子虫在进化树上聚为一支,同源性较高。【结论】东方蜜蜂微孢子虫RRDRP是潜在的亲水性蛋白和胞内蛋白,在东方蜜蜂微孢子虫以及其他微孢子虫和真菌中高度保守。【目的】通过对东方蜜蜂微孢子虫和其他物种的Rad18-like重组与DNA修复蛋白(Rad18-like recombination and DNA repair protein, RRDRP)进行系统进化分析,为进一步研究其功能提供理论依据。【方法】利用Protparam、ProtScale和SWISS-model等软件分析RRDRP的理化性质、亲水性和三级结构。利用SignalP 4.1 Server、NetPhos【Objective】The study aimed to clarify the molecular characteristics of Rad18-like recombination and DNA repair protein(RRDRP)and phyletic evolution of RRDRP proteins from Nosema ceranae and other species, and to provide the theoretical basis for its further functional study. 【Method】Protparam, ProtScale and SWISS-model software were used to investigate the physicochemical property, hydrophilia and tertiary structure of RRDRP in N.ceranae, together with the prediction of the signal peptide, phosphorylation site, transmembrane domain and secondary structure of RRDRP by SignalP 4.1 Server, NetPhos 3.1 Server, TMHMM and SOPMA software. Then the subcellular localization of RRDRP was performed by PSORT software, followed by the prediction of the conserved motifs in RRDRP from N.ceranae and other species by MEME software. Lastly, Mega 11.0 software was employed to perform multiple alignment of amino acid sequences of RRDRP in N.ceranae and other species, prior to the construction of phylogenetic tree of N.ceranae and other 11 species based on RRDRP by neighbor-joining method. 【Result】The RRDRP gene of N.ceranae contained 2 928 nucleotides and encoded 975 amino acids. The molecular weight, lipid solubility coefficient, isoelectric point of RRDRP were 11.50 ku, 93.58 and 6.63, respectively, and it had a molecular formula of C5135H8253N1377O1545S34 and average hydrophilia of -0.627. RRDRP was simultaneously located in nucleus, cytoplasm, peroxisome and cytoskeleton. It consisted of 55 phosphorylation sites, 678 α-helix, 99 extension, 22 β-turn and 176 random coil, but not contained transmembrane domain and signal peptide. There were 5 conserved motifs(Motif 1, Motif 2, Motif 3, Motif 4 and Motif 5)in RRDRP from 11 species including N.ceranae, Encephalitozoon cuniculi and Nosema granulosis, and phylogenetic analysis indicated the closest evolutionary distance between RRDRP from N.ceranae and N.granulosis. 【Conclusion】It was revealed that N.ceranae RRDRP is a potential hydrophilic and intracellular protein,

关 键 词：东方蜜蜂微孢子虫 染色体结构维持蛋白 Rad18-like重组和DNA修复蛋白 分子特性 系统进化 

分 类 号：S895[农业科学—特种经济动物饲养]