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RNA-sequencing of the sturgeon Acipenser baeri provides insights into expression dynamics of morphogenic differentiation and developmental regulatory genes in early versus late developmental stages

机译:the鱼Acipenser baeri的RNA测序可洞悉发育早期和晚期的形态发生分化和发育调控基因的表达动态

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Background Acipenser baeri , one of the critically endangered animals on the verge of extinction, is a key species for evolutionary, developmental, physiology and conservation studies and a standout amongst the most important food products worldwide. Though the transcriptome of the early development of A. baeri has been published recently, the transcriptome changes occurring in the transition from embryonic to late stages are still unknown. The aim of this work was to analyze the transcriptomes of embryonic and post-embryonic stages of A. baeri and identify differentially expressed genes (DEGs) and their expression patterns using mRNA collected from specimens at big yolk plug, wide neural plate and 64?day old sturgeon developmental stages for RNA-Seq. Results The paired-end sequencing of the transcriptome of samples of A. baeri collected at two early (big yolk plug (T1, 32?h after fertilization) and wide neural plate formation (T2, 45?h after fertilization)) and one late (T22, 64?day old sturgeon) developmental stages using Illumina Hiseq2000 platform generated 64039846, 64635214 and 75293762 clean paired-end reads for T1, T2 and T22, respectively. After quality control, the sequencing reads were de novo assembled to generate a set of 149,265 unigenes with N50 value of 1277?bp. Functional annotation indicated that a substantial number of these unigenes had significant similarity with proteins in public databases. Differential expression profiling allowed the identification of 2789, 12,819 and 10,824 DEGs from the respective T1 vs. T2, T1 vs. T22 and T2 vs. T22 comparisons. High correlation of DEGs’ features was recorded among early stages while significant divergences were observed when comparing the late stage with early stages. GO and KEGG enrichment analyses revealed the biological processes, cellular component, molecular functions and metabolic pathways associated with identified DEGs. The qRT-PCR performed for candidate genes in specimens confirmed the validity of the RNA-seq data. Conclusions This study presents, for the first time, an extensive overview of RNA-Seq based characterization of the early and post-embryonic developmental transcriptomes of A. baeri and provided 149,265 gene sequences that will be potentially valuable for future molecular and genetic studies in A. baeri.
机译:背景cip是濒临灭绝的极度濒危动物之一,是进行进化,发育,生理和保护研究的关键物种,在全球最重要的食品中脱颖而出。尽管最近已经公开了A. baeri早期发育的转录组,但是从胚胎到晚期的过渡过程中发生的转录组变化仍然未知。这项工作的目的是分析从大卵黄塞,宽神经板和64天的标本收集的mRNA来分析A. baeri的胚胎和胚后阶段的转录组,并鉴定差异表达基因(DEG)及其表达模式。 RNA-Seq的老development鱼发育阶段。结果在两个早期(大卵黄塞(受精后T1,受精后32?h)和宽大的神经板形成(受精后T2,45?h))和一个晚期收集到的拟南芥样品转录组的配对末端测序(T22,64天大的鱼)使用Illumina Hiseq2000平台的发育阶段分别为T1,T2和T22生成了64039846、64635214和75293762干净的配对末端读数。经过质量控制后,从头开始对测序读数进行组装,以生成一组149,265个单基因,N50值为1277bp。功能注释表明大量这些单基因与公共数据库中的蛋白质具有显着相似性。差异表达谱分析允许从各自的T1对T2,T1对T22和T2对T22的比较中鉴定出2789、12819和10824个DEG。 DEGs的特征在早期阶段之间具有高度相关性,而将晚期阶段与早期阶段进行比较时则观察到明显的差异。 GO和KEGG富集分析揭示了与已鉴定DEG相关的生物学过程,细胞成分,分子功能和代谢途径。对样本中候选基因进行的qRT-PCR证实了RNA-seq数据的有效性。结论本研究首次全面介绍了基于RNA-Seq的拟南芥胚芽早期和胚胎后发育转录组的表征,并提供了149,265个基因序列,这些序列可能对未来的分子生物学和遗传学研究具有潜在的价值。贝里

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