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全长转录组分析通过单分子长读测序。

Full-Length Transcriptome Analysis of by Single-Molecule Long-Read Sequencing.

机构信息

Research Center for Translational Medicine, Key Laboratory of Arrhythmias of the Ministry of Education of China, East Hospital, Tongji University School of Medicine, Shanghai, China.

State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.

出版信息

Front Cell Infect Microbiol. 2021 Feb 23;11:631545. doi: 10.3389/fcimb.2021.631545. eCollection 2021.

Abstract

Malaria, an infectious disease caused by parasites, still accounts for amounts of deaths annually in last decades. Despite the significance of as a model organism of malaria parasites, our understanding of gene expression of this parasite remains largely elusive since lots of progress on its genome and transcriptome are based on assembly with short sequencing reads. Herein, we report the new version of transcriptome dataset containing all full-length transcripts over the whole asexual blood stages by adopting a full-length sequencing approach with optimized experimental conditions of cDNA library preparation. We have identified a total of 393 alternative splicing (AS) events, 3,623 long non-coding RNAs (lncRNAs), 1,555 alternative polyadenylation (APA) events, 57 transcription factors (TF), 1,721 fusion transcripts in . Furthermore, the shotgun proteome was performed to validate the full-length transcriptome of . More importantly, integration of full-length transcriptomic and proteomic data identified 160 novel small proteins in lncRNA regions. Collectively, this full-length transcriptome dataset with high quality and accuracy and the shotgun proteome analyses shed light on the complex gene expression in malaria parasites and provide a valuable resource for related functional and mechanistic researches on genes.

摘要

疟疾是一种由寄生虫引起的传染病,在过去几十年中仍有大量死亡病例。尽管 作为疟原虫模型生物具有重要意义,但由于其基因组和转录组的大量进展都是基于短测序reads 的组装,因此我们对这种寄生虫的基因表达仍然知之甚少。在此,我们通过采用全长测序方法并优化 cDNA 文库制备的实验条件,报告了包含整个无性血期全长转录本的新版本转录组数据集。我们总共鉴定了 393 个可变剪接(AS)事件、3623 个长非编码 RNA(lncRNA)、1555 个可变多聚腺苷酸化(APA)事件、57 个转录因子(TF)、1721 个融合转录本。此外,还进行了鸟枪法蛋白质组学分析,以验证 的全长转录组。更重要的是,全长转录组和蛋白质组数据的整合在 lncRNA 区域中鉴定出 160 个新的小蛋白。总之,这个具有高质量和准确性的全长转录组数据集以及鸟枪法蛋白质组学分析为疟原虫复杂的基因表达提供了新的认识,并为 基因的相关功能和机制研究提供了有价值的资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17fe/7942025/a97999df8c7a/fcimb-11-631545-g001.jpg

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