基于高通量测序技术的敖鲁古雅驯鹿鹿茸转录组分析

doi:10.16431/j.cnki.1671-7236.2020.07.002

摘要/Abstract

摘要： 试验旨在通过高通量测序技术获得驯鹿的遗传信息和鹿茸组织的转录组特征，并进一步挖掘其中与鹿茸经济性状相关的关键基因信息。对驯鹿鹿茸组织提取RNA，检测RNA纯度、完整性及是否有污染，合格后构建文库，文库构建成功需要进行库检，库检合格后使用Illuminanovaseq平台进行转录组测序分析。利用Diamond、HMMER、KAAS、Blast2GO等软件对驯鹿鹿茸转录组序列进行了功能注释、基因丰度分析和其他分析。结果显示，测序获得47 818 202条raw reads，经过滤和质量检查得到了46 964 478条clean reads，组装后得到49 171个Unigenes，表明本次测序获得了高质量的鹿茸转录组。基于序列一致性分析，Unigenes与NR、Swiss-Prot、KOG、KEGG、Pfam、GO数据库比对注释成功总共占89.54%，共44 029条序列。Corset聚类后得到49 171个Unigenes，在这些Unigenes中共找到13 795个SNPs位点和18 446个SSRs位点。GO注释结果显示22 955个Unigenes得到注释，占总注释结果的46.68%。与KEGG数据库进行比对分析，发现49 171个Unigenes可能参与或涉及代谢途径，其中20 258个Unigenes共注释到32个KEGG代谢通路。此外，在转录组结果中发现了一些高丰度的基因，如OPN、TMSB4、TMSB10等，它们的功能可能与驯鹿生茸有关。本试验结果不仅对驯鹿的基因组数据进行了补充，而且还初步揭示了生长期驯鹿鹿茸的基因特征，为驯鹿分子遗传学研究、资源保护与利用及种群资源恢复提供了参考。

关键词: 驯鹿; 鹿茸; 高通量测序; 转录组

Abstract: This experiment was aimed to obtain the genetic information of reindeer and the transcriptome characteristics of antler tissue by high-throughput sequencing technology,and further explore the key gene information related to the economic traits of antler.RNA was extracted from antler tissue in reindeer,and then the purity,integrity and contamination of RNA was detected.Qualified RNA was used for library preparation.The library preparation needs to be tested,after the test,transcriptome sequencing analysis of antler in reindeer using the Illuminanovaseq platform.Functional annotation,gene abundance analysis and other analysis of the transcriptome sequences of antler in reindeer were performed using software such as Diamond,HMMER,KAAS and Blast2GO.The results showed that 47 818 202 raw reads were obtained by sequencing,46 964 478 clean reads were obtained after filtering and quality control,and 49 171 Unigenes were obtained after assembly,indicating that the high-quality antler transcriptome was obtained by sequencing.Based on sequence identity analysis,Unigenes were compared with NR,Swiss-Prot,KOG,KEGG,Pfam and GO databases,the annotations accounted for 89.54% successfully,with a total of 44 029 sequences.After clustering of Corset,49 171 Unigenes were obtained,and 13 795 SNPs and 18 446 SSRs were found in Unigenes.22 955 Unigenes annotated with GO terms,accouting for 46.68% of the total annotated results.Compared with the KEGG database,49 171 Unigenes might be involved in the metabolic pathway,and 20 285 Unigenes were annotated to 32 KEGG metabolic pathways.In addition,there were some high-abundance genes in transcriptome results,such as OPN, TMSB4 and TMSB10.Their function might be related to anlter in reindeer.In summary,this results not only supplement the genomic data of reindeer,but also reveal the genetic characteristics of antler in reindeer in the growing season,which provided a theoretical basis for molecular genetics research,resource conservation and utilization and restoration of population resources.

Key words: reindeer; antler; high-throughput sequencing; transcriptome

中图分类号:

鞠妍, 刘华淼, 何金明, 王磊, 赵佩, 张然然, 董依萌, 董世武, 唐丽昕, 邢秀梅. 基于高通量测序技术的敖鲁古雅驯鹿鹿茸转录组分析[J]. 中国畜牧兽医, 2020, 47(7): 1981-1989.

JU Yan, LIU Huamiao, HE Jinming, WANG Lei, ZHAO Pei, ZHANG Ranran, DONG Yimeng, DONG Shiwu, TANG Lixin, XING Xiumei. Transcriptome Analysis of Antler in Aoluguya Reindeer Based on High-throughput Sequencing Technology[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 1981-1989.

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