本文選題：斯氏副柔線蟲 + 轉(zhuǎn)錄組��； 參考：《中國(guó)農(nóng)業(yè)科學(xué)》2017年23期

【摘要】：【目的】探明不同發(fā)育階段駱駝斯氏副柔線蟲的轉(zhuǎn)錄組差異,了解不同發(fā)育階段蟲體在功能分類和代謝通路等方面的生物學(xué)特征,挖掘生長(zhǎng)發(fā)育相關(guān)功能基因,豐富寄生性線蟲轉(zhuǎn)錄組學(xué)信息�！痉椒ā坎捎肐llumina Hi Seq2000TM高通量測(cè)序技術(shù)對(duì)斯氏副柔線蟲蟲卵、第三期幼蟲和雌蟲進(jìn)行轉(zhuǎn)錄組測(cè)序,構(gòu)建3個(gè)樣本的c DNA文庫,評(píng)估建庫質(zhì)量;利用Trinity軟件對(duì)所得序列進(jìn)行De novo組裝及組裝效率評(píng)估;之后對(duì)獲得的有效序列進(jìn)行功能注釋及相關(guān)生物信息學(xué)分析�！窘Y(jié)果】測(cè)序及組裝后蟲卵、第三期幼蟲和雌蟲分別獲得47 717、76 342和54 624個(gè)Unigenes,在蟲卵和第三期幼蟲比對(duì)中共有33 579個(gè)差異表達(dá)基因,其中表達(dá)上調(diào)的基因有20 477個(gè),表達(dá)下調(diào)的基因有13 102個(gè);在雌蟲和第三期幼蟲比對(duì)中共有32 199個(gè)差異表達(dá)基因,其中表達(dá)上調(diào)的基因有9 293個(gè),表達(dá)下調(diào)的基因有22 906個(gè)。將成對(duì)比較的差異表達(dá)基因分別進(jìn)行GO功能分類,其中蟲卵和第三期幼蟲比對(duì)中有6 617、3 891和8 755個(gè)Unigenes,雌蟲和第三期幼蟲比對(duì)中有7 043、3 686和10 177個(gè)Unigenes分別注釋到生物過程、細(xì)胞組成和分子功能三大類中;通過KEGG pathway數(shù)據(jù)庫分析,蟲卵和第三期幼蟲比對(duì)中有6 521個(gè)差異表達(dá)基因參與到251條通路中,顯著富集MAPK信號(hào)通路、Wnt信號(hào)通路和氧化磷酸化等通路,雌蟲和第三期幼蟲比對(duì)中有6 528個(gè)參與到251條通路中,顯著富集新陳代謝通路、DNA復(fù)制和細(xì)胞周期等通路。差異表達(dá)基因功能聚類分析中,雌蟲和蟲卵在調(diào)控生長(zhǎng)速率和生殖發(fā)育等方面高表達(dá);第三期幼蟲在防御機(jī)制和糖類代謝等方面高表達(dá);且3個(gè)發(fā)育階段均在胚胎發(fā)育,胚后發(fā)育中富集高表達(dá),其中在胚胎發(fā)育和胚后發(fā)育中分別有242和202個(gè)相關(guān)功能基因在3個(gè)發(fā)育階段都表達(dá),這些基因可能在胚胎(后)發(fā)育過程中起核心作用。此外,獲得了9種異時(shí)性基因(如:LIN-28、LIN-14和RHEB-1等)、48個(gè)核激素受體(NHRs),包括NHR-49,NHR-48,NHR-40,NHR-1等以及36個(gè)鋅金屬蛋白酶(NAS),包括NAS-36、NAS-33和NAS-14等,并對(duì)其在斯氏副柔線蟲蟲卵、第三期幼蟲和雌蟲中的富集程度進(jìn)行分析,發(fā)現(xiàn)這些基因在維持線蟲正常生長(zhǎng)發(fā)育過程中發(fā)揮關(guān)鍵作用,【結(jié)論】利用RNA-seq技術(shù)對(duì)3個(gè)發(fā)育階段的斯氏副柔線蟲進(jìn)行測(cè)序和生物信息學(xué)分析,研究了不同發(fā)育階段蟲體的差異表達(dá)基因在GO功能分類、KEGG代謝通路和基因功能聚類等方面的生物學(xué)特性,篩選出多種異時(shí)性基因和發(fā)育相關(guān)重要基因,為后續(xù)開展斯氏副柔線蟲功能基因組學(xué)研究,蟲體與宿主互作、致病機(jī)制、免疫逃避等研究提供了理論依據(jù)。
[Abstract]:[Objective] to explore the differences in the transcriptional groups of camel's parasitic nematode in different developmental stages, to understand the biological characteristics of different developmental stages in functional classification and metabolic pathways, to excavate growth and development related genes and to enrich the transcriptional information of parasitic nematodes. [Methods] high throughput sequencing of Illumina Hi Seq2000TM was used. Technology was used to sequence the egg, third stage larvae and females of the Coriolis slovaki, construct the C DNA Library of 3 samples, evaluate the quality of the building, and evaluate the assembly and assembly efficiency of the De novo using Trinity software; and then the effective sequence was annotated and the related bioinformatics analysis. The third stage larvae and females obtained 47 717,76 342 and 54624 Unigenes respectively. There were 33579 differentially expressed genes in the egg and third stage larvae, of which 20477 genes were up-regulated, and 13102 expressed down regulated genes, and 32199 differences were expressed in the female and third stage larvae. In the gene, there were 9293 up-regulated genes and 22906 down-regulated genes. The differentially expressed genes were divided into GO functional classifications, including 6 617,3 891 and 8755 Unigenes for insect eggs and third stage larvae, and 7 043,3 686 and 10177 Unigenes respectively annotated by 7 043,3 686 and 10177 Unigenes respectively. In the three major categories of biological processes, cell composition and molecular function, through the analysis of KEGG pathway database, 6521 differentially expressed genes in the egg and third stage larvae were involved in the 251 pathways, which significantly enriched the MAPK signaling pathway, the Wnt signaling pathway and the oxidative phosphorylation pathway, and 6528 of the third and third stage larvae were involved. In the 251 pathways, the metabolic pathways, DNA replication and cell cycle pathways were significantly enriched. In the functional cluster analysis of differentially expressed genes, the female and the eggs were highly expressed in the regulation of growth rate and reproductive development, and the third stage larvae were highly expressed in the defense mechanism and carbohydrate metabolism, and the 3 developmental stages were in the embryonic development and after the embryo. There are 242 and 202 related functional genes in the embryonic development and postembryonic development, respectively, in which 242 and 202 related genes are expressed in 3 developmental stages. These genes may play a core role in the development of the embryo. In addition, 9 kinds of isochronic genes (such as LIN-28, LIN-14 and RHEB-1, etc.), and 48 nuclear hormone receptors (NHRs), including NHR-, are obtained. 49, NHR-48, NHR-40, NHR-1 and 36 zinc metalloproteinases (NAS), including NAS-36, NAS-33 and NAS-14, and their enrichment in the eggs of the Coriolis slovaki, third stage larvae and females, and found that these genes play a key role in maintaining normal growth and development of the nematodes. [Conclusion] 3 with RNA-seq technology is used. By sequencing and bioinformatics analysis, the biological characteristics of differential expression genes of different developmental stages of insect body in GO function classification, KEGG metabolic pathway and gene function clustering were studied. The research on insect functional genomics provides a theoretical basis for the interaction between host and host, pathogenesis and immune evasion.
【作者單位】： 內(nèi)蒙古農(nóng)業(yè)大學(xué)獸醫(yī)學(xué)院/農(nóng)業(yè)部動(dòng)物疾病臨床診療技術(shù)重點(diǎn)實(shí)驗(yàn)室;內(nèi)蒙古農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(31260603)
【分類號(hào)】：S852.731
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本文編號(hào)：1942813