文章摘要
日本医蛭唾液腺对饥饿胁迫响应的转录组比较分析
Comparative transcriptome analysis of salivary glands of Hirudo nipponia in response to starvation
投稿时间:2018-12-19  修订日期:2019-10-13
DOI:
中文关键词: 日本医蛭  唾液腺  饥饿胁迫  转录组测序  从头组装  差异表达分析  
英文关键词: Hirudo nipponia  salivary glands  starvation stress  transcriptome sequence  De novo assembly  differential expression analysis
基金项目:国家自然科学基金(31440030)
作者单位E-mail
邢月婷 中国计量大学生命科学学院 690748736@qq.com 
管峰 中国计量大学生命科学学院  
李诗语 中国计量大学生命科学学院  
张世雄 中国计量大学生命科学学院  
罗媛媛 中国计量大学生命科学学院 yyluo@cjlu.edu.cn 
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中文摘要:
      [目的]为了探究饥饿胁迫对日本医蛭唾液腺基因表达水平的影响;[方法]本实验通过Illumina Hiseq 2500高通量测序平台分别对饥饿30天处理组(D30)和饥饿0天(D0)对照组的日本医蛭的唾液腺组织进行双端测序;[结果]对获得的原始数据进行质量控制及从头组装,获得145981个unigenes,平均长度为675bp,N50为1127bp。针对CDD、KOG、COG、NR、NT、PFAM、Swissprot、TrEMBL、GO和KEGG等数据库的序列比对分析,145981个unigenes均能得到注释。通过引入TPM来估算基因表达水平,并通过DEGseq进行基因表达差异分析。以饥饿0天为对照组,显著差异基因筛选条件设置为Q Value<0.05且差异倍数|FoldChange|>2,获得2650个差异基因,其中667个基因显著上调,1983个基因显著下调。选取4个日本医蛭唾液腺重要功能基因进行RT-qPCR验证,结果证明转录组测序分析可靠。将所有差异表达基因进行GO功能富集分析,显著富集到175条途径,其中胞质核糖体途径富集程度最为显著,核糖体途径次之,且参与这些途径的差异基因基本均下调。KEGG通路富集分析进一步证明,差异基因在核糖体通路中富集程度最为显著。此外,差异基因中4个基因被预测参与抗凝,抗血栓,抗菌,抗炎和抗肿瘤过程,这可能在各种疾病的治疗中发挥重要作用;[结论]参与核糖体通路的基因表达显著降低,表明日本医蛭唾液腺通过降低蛋白质代谢以应对饥饿环境。该研究结果为下一步深入研究日本医蛭唾液腺饥饿胁迫适应的分泌调控机制以及药用价值基因的发掘提供了重要的参考材料,并为其他医学蛭类研究提供参考依据。
英文摘要:
      In order to investigate the variations of gene expression of salivary gland in Hirudo nipponia under starvation stress, two libraries of starvation treatment group (D30) and control group(D0) were constructed and sequenced using the Illumina HiSeq 2500 platform (paired-end). After stringent quality control of raw data, 145981 unigenes were obtained using de novo assembly. The average length of unigenes was 675 bp and N50 lengths was 1127 bp. And then all of unigenes were annotated via sequence alignment analysis in CDD, KOG, COG, NR, NT, PFAM, Swissprot, TrEMBL, GO and KEGG databases. The amount of gene expression was also estimated according to TPM (Transcripts Per Kilobase Million). By comparing the transcriptome data of D30 sample and D0 sample, the differentially expressed genes (DEGs) were screened out with a threshold criteria Q Value<0.05 and |FoldChange|> 2. Totally 2650 DEGs were identified under starvation stress, among which 667 genes were up-regulated and 1983 genes were down-regulated. Four functional DEGs were selected for RT-qPCR analysis, and the results confirmed that the transcriptome analysis was reliable. In addition, 175 pathways were significantly enriched when the DEGs were subject to GO enrichment analysis. The results showed that most of the DEGs were involved in cytoplasmic ribosome pathway and ribosome pathway. The DEGs were almost down-regulated in these pathways. It also was proved in KEGG enrichment analysis that most DEGs were involved in ribosome pathway. The DEGs down-regulated in ribosome pathway can indicate that it reduces protein metabolism under starvation stress. Futhermore, were predicted to be involved in anticoagulatory, antithrombotic, antibacterial, anti-inflammatory and antitumor processes, which might play important roles in the treatment of various diseases. The results above mentioned provide foundation information for further study on the molecular mechanism of starvation tolerance of salivary gland and the discovery of pharmaceutical value genes in Hirudo nipponia, and also provides reference for other medicinal leeches.
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