作者：孙明会,钱加珺,李琪明,张天泽,张佳欣,赵浩东,张奎昊,朱乐冉,陈田,陈大福,付中民,郭睿
单位：1 福建农林大学动物科学学院(蜂学学院)，福建 福州 350002,2 福建农林大学蜂疗研究所，福建 福州 350002
关键词：微孢子虫,东方蜜蜂微孢子虫,RNA甲基化,N6-甲基腺苷,甲基转移酶,侵染,增殖
基金：国家自然科学基金面上项目(32172792),国家现代农业产业技术体系专项资金(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,福建农林大学动物科学学院(蜂学学院),国家级大学生创新创业训练计划项目(202110389027),福建省大学生创新创业训练计划(202210389115),福建省大学生创新创业训练计划(202210389128)
N6-甲基腺苷(N6-methyladenosine, m6A)是真核生物中分布广泛、含量丰富且作用关键的一类RNA修饰。东方蜜蜂微孢子虫Nosema ceranae专性侵染成年蜜蜂导致微孢子病，给养蜂业造成很大损失。本研究对东方蜜蜂微孢子虫的甲基转移酶样蛋白5 (Methyltransferase-like protein 5, Mettl5)基因NcMettl5进行克隆，通过生物信息学预测NcMettl5蛋白的理化性质和分子特性，并通过RT-qPCR检测NcMettl5在东方蜜蜂微孢子虫侵染意大利蜜蜂Apis mellifera ligustica工蜂过程的相对表达量，以期丰富NcMettl5的信息，并为探究东方蜜蜂微孢子侵染中NcMettl5的功能及调控机制提供基础。结果表明，扩增出约600 bp的目的片段，与GenBank数据库收录的预测出的NcMettl5序列一致；NcMettl5的分子量约为22.64 kDa，分子式为C1039H1645N269O284S6，理论等电点是9.37，脂溶系数是98.98，不稳定系数为13.96；包含196个氨基酸和20个磷酸化位点；平均亲水系数为?0.230，不含典型的跨膜结构域和信号肽，可同时定位于细胞骨架、细胞质和细胞核、细胞核、线粒体和过氧化物酶体；NcMettl5含有1个超家族结构域：AdoMet_MTases superfamily；东方蜜蜂微孢子虫、家蚕微孢子虫Nosema bombycis CQ1和颗粒病微孢子虫Nosema granulosis的Mettl5均含有7个相同的保守基序，序列一致性较高，且进化距离较近；NcMettl5在东方蜜蜂微孢子虫接种后1-4 d的意大利蜜蜂工蜂中肠内均有表达；相较于接种后1 d (1 day post inoculation, 1 dpi)，NcMettl5的表达量在2 dpi和3 dpi下调但差异不显著(P>0.05)，在4 dpi显著下调(P<0.05)。研究结果表明成功克隆到东方蜜蜂微孢子虫的NcMettl5基因，明确了NcMettl5蛋白的分子特性，并揭示NcMettl5可能为亲水性蛋白、非跨膜蛋白和胞内蛋白，Nosema属的东方蜜蜂微孢子虫、家蚕微孢子虫和颗粒病微孢子虫的Mettl5蛋白具有较高的保守性，NcMettl5在东方蜜蜂微孢子虫增殖周期内真实表达且总体呈持续下降的表达趋势。"/>template{display:none;}.mag-rich-xref-fn { cursor: pointer;}//长视频$(document).ready(function(){if($("#showLongArticleVideo") && $("#showLongArticleVideo").length && $("#showLongArticleVideo").length>0){mag_ajax_update({ele_id:'showLongArticleVideo',url:mag_currentQikanUrl() + "/CN/article/showLongArticleVideo.do?id="+$("#articleId").val()});}});//可视化图谱$(document).ready(function(){if($("#showArticle_knowledgeMap") && $("#showArticle_knowledgeMap").length && $("#showArticle_knowledgeMap").length>0){jQuery.getJSON("/Jwxb/CN/lexeme/existsCnctstInArticle.do?articleID="+$("#articleId").val(), null, function(json){if(json.result){$("#showArticle_knowledgeMap").show();}});}});$(document).ready(function(){var metaDataStr = $.trim($("#article_metaJson").val());if(metaDataStr!=''){eval("window.metaData="+metaDataStr);}if(window.metaData && (!window.metaData.authorNotesCommon_cn && !window.metaData.authorNotesCorresp_cn && window.metaData.authorNotes_cn)){window.metaData.authorNotesCommon_cn = window.metaData.authorNotes_cn;}if(window.metaData && (!window.metaData.authorNotesCommon_en && !window.metaData.authorNotesCorresp_en && window.metaData.authorNotes_en)){window.metaData.authorNotesCommon_en = window.metaData.authorNotes_en;}var _nlmdtdXml = $("#article_nlmdtdXml").val();var magDir = _nlmdtdXml.replace(/\/[^\/]+$/,'');window.metaData.magDir = magDir;new mag_vue({ el: '#metaVue', //dataUrl: mag_currentQikanUrl() + '/EN/article/getRichHtmlJson.do?articleId='+$("#articleId").val(), data:window.metaData, loading:true, doTextFun:function(text){ return doDataJsonText(text); 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来源：2023年第5期
《菌物学报》期刊编辑部
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