本文主要研究内容
作者丁艳娟,刘永康,罗雨嘉,邓颖梅,徐红星,唐斌,徐彩娣（2019）在《褐飞虱GSK-3调控糖原与海藻糖代谢的潜在功能》一文中研究指出：【目的】昆虫胰岛素信号途径能够介导糖原合成酶激酶3(glycogen synthase kinase 3,简称GSK-3或GSK3)调控体内糖原及海藻糖等糖代谢过程,从而控制昆虫的各项生命活动。论文旨在探究糖原合成酶激酶在褐飞虱(Nilaparvata lugens)体内对糖原与海藻糖代谢的调控作用。【方法】首先,基于GSK-3的cDNA编码序列,利用ExPASy工具翻译GSK-3氨基酸序列,预测蛋白分子量大小及等电点(pI);然后利用SignaIP4.1Server对其信号肽进行分析。其次,以笔者实验室饲养的褐飞虱为研究对象,从4龄开始,每12 h取材,取至成虫48 h。利用Trizol法提取褐飞虱总RNA,根据反转录试剂盒合成第一链DNA,以18S作为内参基因,通过实时荧光定量PCR(qRT-PCR)检测褐飞虱GSK-3在不同龄期mRNA水平上的相对表达量。然后利用RNAi技术,向褐飞虱体内显微注射双链RNA(dsRNA)抑制GSK-3,以注射ds GFP的褐飞虱作为对照组。注射后48 h利用qRT-PCR技术检测GSK-3的表达情况,确定抑制效果。另外,取注射后48 h虫体,分别测定褐飞虱体内海藻糖、葡萄糖、糖原含量及海藻糖酶(trehalase,TRE)活性变化。最后采用qRT-PCR检测胰岛素信号通路胰岛素受体基因(insulin receptor,InR)、类胰岛素多肽基因(insulin-like peptides,Ilps)及海藻糖代谢途径TRE、海藻糖合成酶基因(trehalose-6-phophate synthase,TPS)、糖原磷酸化酶基因(glycogen phosphorylase,GP)、糖原合成酶基因(glycogen synthase,GS)中相关基因的表达,分析GSK-3在胰岛素信号通路及海藻糖代谢途径中的调控作用。【结果】褐飞虱GSK-3开放阅读框为1 914 bp,编码637个氨基酸;预测蛋白分子量为69.25 kD,等电点为9.15,为偏碱性蛋白,无信号肽结构,序列高度保守。发育表达模式结果显示GSK-3在不同发育阶段表达不一致,5龄若虫蜕皮前后低表达。GSK-3的dsRNA注射后48 h,与对照组ds GFP相比,GSK-3表达极显著下降,表明RNA干扰效果明显。糖原含量和两类海藻糖酶活性显著下降,而海藻糖含量显著上升,推测糖原和葡萄糖转化为海藻糖,作为其生理活动的能量来源。qRT-PCR检测发现,当GSK-3表达抑制后48 h,TRE1-2的表达量显著下降,而TRE1-1和TRE2的表达量极显著下降。另外,2个TPS基因、GS以及GP的表达量均极显著下降;胰岛素信号通路的2个InR基因和4个Ilps基因的表达同样被抑制,间接表明InR能够调控GSK-3的表达。【结论】褐飞虱GSK-3低表达后能够通过调控胰岛素信号通路及海藻糖代谢途径相关基因表达来调控糖原及海藻糖代谢。相关研究结果有助于更加全面地探索褐飞虱等昆虫糖原合成酶激酶调控海藻糖及糖类物质平衡的潜在分子机理。
Abstract
【mu de 】kun chong yi dao su xin hao tu jing neng gou jie dao tang yuan ge cheng mei ji mei 3(glycogen synthase kinase 3,jian chen GSK-3huo GSK3)diao kong ti nei tang yuan ji hai zao tang deng tang dai xie guo cheng ,cong er kong zhi kun chong de ge xiang sheng ming huo dong 。lun wen zhi zai tan jiu tang yuan ge cheng mei ji mei zai he fei shi (Nilaparvata lugens)ti nei dui tang yuan yu hai zao tang dai xie de diao kong zuo yong 。【fang fa 】shou xian ,ji yu GSK-3de cDNAbian ma xu lie ,li yong ExPASygong ju fan yi GSK-3an ji suan xu lie ,yu ce dan bai fen zi liang da xiao ji deng dian dian (pI);ran hou li yong SignaIP4.1Serverdui ji xin hao tai jin hang fen xi 。ji ci ,yi bi zhe shi yan shi si yang de he fei shi wei yan jiu dui xiang ,cong 4ling kai shi ,mei 12 hqu cai ,qu zhi cheng chong 48 h。li yong Trizolfa di qu he fei shi zong RNA,gen ju fan zhuai lu shi ji he ge cheng di yi lian DNA,yi 18Szuo wei nei can ji yin ,tong guo shi shi ying guang ding liang PCR(qRT-PCR)jian ce he fei shi GSK-3zai bu tong ling ji mRNAshui ping shang de xiang dui biao da liang 。ran hou li yong RNAiji shu ,xiang he fei shi ti nei xian wei zhu she shuang lian RNA(dsRNA)yi zhi GSK-3,yi zhu she ds GFPde he fei shi zuo wei dui zhao zu 。zhu she hou 48 hli yong qRT-PCRji shu jian ce GSK-3de biao da qing kuang ,que ding yi zhi xiao guo 。ling wai ,qu zhu she hou 48 hchong ti ,fen bie ce ding he fei shi ti nei hai zao tang 、pu tao tang 、tang yuan han liang ji hai zao tang mei (trehalase,TRE)huo xing bian hua 。zui hou cai yong qRT-PCRjian ce yi dao su xin hao tong lu yi dao su shou ti ji yin (insulin receptor,InR)、lei yi dao su duo tai ji yin (insulin-like peptides,Ilps)ji hai zao tang dai xie tu jing TRE、hai zao tang ge cheng mei ji yin (trehalose-6-phophate synthase,TPS)、tang yuan lin suan hua mei ji yin (glycogen phosphorylase,GP)、tang yuan ge cheng mei ji yin (glycogen synthase,GS)zhong xiang guan ji yin de biao da ,fen xi GSK-3zai yi dao su xin hao tong lu ji hai zao tang dai xie tu jing zhong de diao kong zuo yong 。【jie guo 】he fei shi GSK-3kai fang yue dou kuang wei 1 914 bp,bian ma 637ge an ji suan ;yu ce dan bai fen zi liang wei 69.25 kD,deng dian dian wei 9.15,wei pian jian xing dan bai ,mo xin hao tai jie gou ,xu lie gao du bao shou 。fa yo biao da mo shi jie guo xian shi GSK-3zai bu tong fa yo jie duan biao da bu yi zhi ,5ling re chong tui pi qian hou di biao da 。GSK-3de dsRNAzhu she hou 48 h,yu dui zhao zu ds GFPxiang bi ,GSK-3biao da ji xian zhe xia jiang ,biao ming RNAgan rao xiao guo ming xian 。tang yuan han liang he liang lei hai zao tang mei huo xing xian zhe xia jiang ,er hai zao tang han liang xian zhe shang sheng ,tui ce tang yuan he pu tao tang zhuai hua wei hai zao tang ,zuo wei ji sheng li huo dong de neng liang lai yuan 。qRT-PCRjian ce fa xian ,dang GSK-3biao da yi zhi hou 48 h,TRE1-2de biao da liang xian zhe xia jiang ,er TRE1-1he TRE2de biao da liang ji xian zhe xia jiang 。ling wai ,2ge TPSji yin 、GSyi ji GPde biao da liang jun ji xian zhe xia jiang ;yi dao su xin hao tong lu de 2ge InRji yin he 4ge Ilpsji yin de biao da tong yang bei yi zhi ,jian jie biao ming InRneng gou diao kong GSK-3de biao da 。【jie lun 】he fei shi GSK-3di biao da hou neng gou tong guo diao kong yi dao su xin hao tong lu ji hai zao tang dai xie tu jing xiang guan ji yin biao da lai diao kong tang yuan ji hai zao tang dai xie 。xiang guan yan jiu jie guo you zhu yu geng jia quan mian de tan suo he fei shi deng kun chong tang yuan ge cheng mei ji mei diao kong hai zao tang ji tang lei wu zhi ping heng de qian zai fen zi ji li 。
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论文详细介绍
论文作者分别是来自中国农业科学的丁艳娟,刘永康,罗雨嘉,邓颖梅,徐红星,唐斌,徐彩娣，发表于刊物中国农业科学2019年07期论文，是一篇关于褐飞虱论文,干扰论文,糖原合成酶激酶论文,糖原与海藻糖代谢论文,实时荧光定量论文,中国农业科学2019年07期论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自中国农业科学2019年07期论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：褐飞虱论文; 干扰论文; 糖原合成酶激酶论文; 糖原与海藻糖代谢论文; 实时荧光定量论文; 中国农业科学2019年07期论文;