本文主要研究内容
作者黄敏婕（2019）在《长白猪和金华猪肝脏组织mRNA、microRNA和circRNA的比较研究》一文中研究指出：环状RNA(circular RNA,circRNA)是一类没有5’末端帽子与3’末端poly A尾巴结构的闭合环状非编码RNA分子,可作为内源性竞争RNA(ceRNA)且能够顺式调控亲本基因的表达。研究发现肝脏脂质代谢受到非编码RNA介导的转录前后多方面的调控。目前对于肝脏中circRNA的表达与调控研究相对匮乏。本研究选取70日龄未去势的长白猪与金华猪公猪为研究对象,比较两个品种的血清生化水平,利用高通量测序技术对两者的肝脏组织进行mRNA、miRNA、circRNA的测序研究,筛选出差异表达的基因、miRNA、circRNA,并通过荧光定量PCR技术对测序结果进行了验证,对差异表达的基因、差异表达miRNA的靶基因和差异表达circRNA的亲本基因进行功能注释和通路分析,此外还对差异miRNA与差异基因进行了联合分析、circRNA与miRNA的互作网络进行了预测分析。主要研究结果如下:(1)长白猪体重和肝重均极显著高于金华猪(P<0.01),但两者的肝重/体重比值无差异。长白猪的血清胰岛素、TT3、TT4含量显著高于金华猪(P<0.05),金华猪的血清TCH和LDL-C极显著高于长白猪(P<0.01),长白猪的血糖和HDL-C水平高于金华猪,但差异并不显著。(2)通过DGE测序获得共约78 M(million)读长为50 nt的单末端Clean reads,超过86.64%能够比对到参考基因组上。差异表达分析筛选到差异基因467个(|log2(foldchange)|>1,P-adjust＜0.05,下同),其中 172个基因在金华猪中上调表达。差异基因GO富集KEGG通路分析揭示它们主要涉及有氧化还原、脂肪生物合成、细胞脂质代谢等生物学过程,并与细胞外基质受体相互作用、蛋白质消化与吸收、脂肪酸代谢、PPAR信号通路和脂肪消化和吸收等通路相关。(3)通过小RNA测序共获得约75.5 M的clean reads,超过98.96%能够比对到参考基因组上。在两个品种的肝脏中共鉴定到266个已知miRNA,其中14个miRNA在长白猪中特异性表达,7个miRNA在金华猪中特异性表达,特异性表达的miRNA表达量均较低。差异分析筛选出35个差异表达miRNA,其中8个miRNA在金华猪中上调表达。生物信息学分析发现差异miRNA涉及如内吞作用、脂肪酸生物合成、脂肪酸代谢通路、PI3K-Akt信号通路、TGF-β信号通路、cAMP信号通路等脂质代谢相关的通路。(4)通过circRNA文库测序分析,共鉴定出circRNA 61858个,大部分circRNA来源于外显子,约93%的circRNA呈低表达水平(RPM值<100)。差异分析筛选到376个差异circRNA,有116个circRNA在金华猪中上调表达。差异circRNA的亲本基因富集分析发现它们主要涉及脂肪代谢过程和氧化还原过程中,通过与数字基因表达谱数据对比,发现这些亲本基因在两个品种中呈高表达。(5)随机选取的6个差异基因、6个差异miRNA和8个差异circRNA经qRT-PCR检测发现,它们在长白猪和金华猪肝脏中的表达量与高通量测序结果基本一致。(6)对差异miRNA、mRNA和circRNA进行联合分析,结果得到的PPAR通路相关调控网络图中包括10个mRNA、11个miRNA和32个circRNA。
Abstract
huan zhuang RNA(circular RNA,circRNA)shi yi lei mei you 5’mo duan mao zi yu 3’mo duan poly Awei ba jie gou de bi ge huan zhuang fei bian ma RNAfen zi ,ke zuo wei nei yuan xing jing zheng RNA(ceRNA)ju neng gou shun shi diao kong qin ben ji yin de biao da 。yan jiu fa xian gan zang zhi zhi dai xie shou dao fei bian ma RNAjie dao de zhuai lu qian hou duo fang mian de diao kong 。mu qian dui yu gan zang zhong circRNAde biao da yu diao kong yan jiu xiang dui kui fa 。ben yan jiu shua qu 70ri ling wei qu shi de chang bai zhu yu jin hua zhu gong zhu wei yan jiu dui xiang ,bi jiao liang ge pin chong de xie qing sheng hua shui ping ,li yong gao tong liang ce xu ji shu dui liang zhe de gan zang zu zhi jin hang mRNA、miRNA、circRNAde ce xu yan jiu ,shai shua chu cha yi biao da de ji yin 、miRNA、circRNA,bing tong guo ying guang ding liang PCRji shu dui ce xu jie guo jin hang le yan zheng ,dui cha yi biao da de ji yin 、cha yi biao da miRNAde ba ji yin he cha yi biao da circRNAde qin ben ji yin jin hang gong neng zhu shi he tong lu fen xi ,ci wai hai dui cha yi miRNAyu cha yi ji yin jin hang le lian ge fen xi 、circRNAyu miRNAde hu zuo wang lao jin hang le yu ce fen xi 。zhu yao yan jiu jie guo ru xia :(1)chang bai zhu ti chong he gan chong jun ji xian zhe gao yu jin hua zhu (P<0.01),dan liang zhe de gan chong /ti chong bi zhi mo cha yi 。chang bai zhu de xie qing yi dao su 、TT3、TT4han liang xian zhe gao yu jin hua zhu (P<0.05),jin hua zhu de xie qing TCHhe LDL-Cji xian zhe gao yu chang bai zhu (P<0.01),chang bai zhu de xie tang he HDL-Cshui ping gao yu jin hua zhu ,dan cha yi bing bu xian zhe 。(2)tong guo DGEce xu huo de gong yao 78 M(million)dou chang wei 50 ntde chan mo duan Clean reads,chao guo 86.64%neng gou bi dui dao can kao ji yin zu shang 。cha yi biao da fen xi shai shua dao cha yi ji yin 467ge (|log2(foldchange)|>1,P-adjust＜0.05,xia tong ),ji zhong 172ge ji yin zai jin hua zhu zhong shang diao biao da 。cha yi ji yin GOfu ji KEGGtong lu fen xi jie shi ta men zhu yao she ji you yang hua hai yuan 、zhi fang sheng wu ge cheng 、xi bao zhi zhi dai xie deng sheng wu xue guo cheng ,bing yu xi bao wai ji zhi shou ti xiang hu zuo yong 、dan bai zhi xiao hua yu xi shou 、zhi fang suan dai xie 、PPARxin hao tong lu he zhi fang xiao hua he xi shou deng tong lu xiang guan 。(3)tong guo xiao RNAce xu gong huo de yao 75.5 Mde clean reads,chao guo 98.96%neng gou bi dui dao can kao ji yin zu shang 。zai liang ge pin chong de gan zang zhong gong jian ding dao 266ge yi zhi miRNA,ji zhong 14ge miRNAzai chang bai zhu zhong te yi xing biao da ,7ge miRNAzai jin hua zhu zhong te yi xing biao da ,te yi xing biao da de miRNAbiao da liang jun jiao di 。cha yi fen xi shai shua chu 35ge cha yi biao da miRNA,ji zhong 8ge miRNAzai jin hua zhu zhong shang diao biao da 。sheng wu xin xi xue fen xi fa xian cha yi miRNAshe ji ru nei tun zuo yong 、zhi fang suan sheng wu ge cheng 、zhi fang suan dai xie tong lu 、PI3K-Aktxin hao tong lu 、TGF-βxin hao tong lu 、cAMPxin hao tong lu deng zhi zhi dai xie xiang guan de tong lu 。(4)tong guo circRNAwen ku ce xu fen xi ,gong jian ding chu circRNA 61858ge ,da bu fen circRNAlai yuan yu wai xian zi ,yao 93%de circRNAcheng di biao da shui ping (RPMzhi <100)。cha yi fen xi shai shua dao 376ge cha yi circRNA,you 116ge circRNAzai jin hua zhu zhong shang diao biao da 。cha yi circRNAde qin ben ji yin fu ji fen xi fa xian ta men zhu yao she ji zhi fang dai xie guo cheng he yang hua hai yuan guo cheng zhong ,tong guo yu shu zi ji yin biao da pu shu ju dui bi ,fa xian zhe xie qin ben ji yin zai liang ge pin chong zhong cheng gao biao da 。(5)sui ji shua qu de 6ge cha yi ji yin 、6ge cha yi miRNAhe 8ge cha yi circRNAjing qRT-PCRjian ce fa xian ,ta men zai chang bai zhu he jin hua zhu gan zang zhong de biao da liang yu gao tong liang ce xu jie guo ji ben yi zhi 。(6)dui cha yi miRNA、mRNAhe circRNAjin hang lian ge fen xi ,jie guo de dao de PPARtong lu xiang guan diao kong wang lao tu zhong bao gua 10ge mRNA、11ge miRNAhe 32ge circRNA。
论文参考文献
论文详细介绍
论文作者分别是来自浙江大学的黄敏婕，发表于刊物浙江大学2019-09-16论文，是一篇关于肝脏论文,长白猪论文,金华猪论文,浙江大学2019-09-16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，资料来自浙江大学2019-09-16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：肝脏论文; 长白猪论文; 金华猪论文; 浙江大学2019-09-16论文;