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绒山羊皮肤全长转录组测序及生物信息学分析
李颖,宋峰,苏德其其格,AKONYANI Zaccheaus Pazamilala,乌兰吐雅,吴江鸿
0
(内蒙古民族大学 动物科技学院, 内蒙古 通辽 028000)
摘要:
为能够更精准地得到绒山羊皮肤mRNA全长序列和完整结构信息,并对新基因和Iso-form进行更全面的分析以对绒山羊转录组数据进行补充,以绒山羊作为试验动物,使用SMRT测序技术,对绒山羊皮肤组织混合样品进行全长转录组测序,并使用TransDecoder、Astalavista和Cytoscape等分析工具对其编码区序列及其对应氨基酸序列、可变剪接类型以及基因功能富集进行预测分析。结果表明:1)测序共获得 42.41 Gb clean data的数据,CCS(Circular consensus sequencing)591 585条,其中全长非嵌合序列 466 058条。2)共鉴定出6 166个新基因,55 875条新转录本,SSR(Simple sequence repeat)66 951个、ORF(Open reading frame)序列39 346条、TF(Transcription factors)5 641个及10 927个lncRNA,并完成49 573条新转录本的功能注释。3)鉴定出25 717种可变剪接,其中内含子保留为主要的剪接方式,基于功能富集分析结果和可变剪接位点信息发现黑素皮质素1受体(Melanocortin 1 receptor,MC1R)与微管蛋白3类(Tubulin-β-III,TUBB3)可发生可变剪接,共用TUBB3第一外显子。鉴定的lncRNA中新lncRNA 8 251个,主要类型为内含子lncRNA。本研究更精准地得到了绒山羊皮肤mRNA全长序列及完整结构信息,为进一步研究绒山羊皮肤基因结构及互作机理奠定基础,同时为绒山羊基因组资源进行了数据补充。
关键词:  绒山羊  SMRT  lncRNA  可变剪接
DOI:10.11841/j.issn.1007-4333.2022.12.14
投稿时间:2022-04-20
基金项目:内蒙古自然科学基金项目(2018MS03034);内蒙古民族大学博士启动基金项目(BS527)
Sequencing and bioinformatics analysis of the full-length transcriptome of cashmere goat skin
LI Ying,SONG Feng,SU Deqiqige,AKONYANI Zaccheaus Pazamilala,WU Lantuya,Wu Jianghong
(College of Animal Science and Technology, Inner Mongolia Minzu University, Tongliao 028000, China)
Abstract:
In order to more accurately obtain the full-length mRNA sequences and complete structural information of cashmere goat skin, and conduct more comprehensive research on new genes and ISO form, and supplement the cashmere goat transcriptome data, cashmere goat was used as an experimental animal in the study. The full-length transcriptome of mixed samples of cashmere goat skin tissue using SMRT sequencing technology as test animals were sequenced. The sequences of coding regions and their corresponding amino acid sequences, variable splice types, and functional enrichment were predicted by using TransDecoder, Astalavista, Cytoscape and other analysis tools. The results showed that: 1)A total of 42. 41 Gb of clean data were obtained by sequencing, with 591 585 CCS(Circular consensus sequencing)sequences, including 466 058 full-length non-chimeric sequences. 2)A total of 6 166 new genes, 55 875 new transcripts, 66 951 SSR(Simple Sequence Repeat), 39 346 ORF(Open Reading Frame)sequences, 5 641 TF(Transcription Factors)and 10 927 lncRNAs were identified, and the functional annotation of 49 573 new transcripts were annotated. 3)Based on the results of functional enrichment analysis and variable splice site information, we found that melanocortin 1 receptor(MC1R)and microtubulin class 3(Tubulin-β-III, TUBB3)could undergo variable splicing and share TUBB3 first exon. There were 8 251 new lncRNAs identified, and the main type was intronic lncRNA. In conclusion, the full-length sequence and complete structural information of the mRNA of the skin of cashmere goat is obtained, which lays the foundation for further study of the gene structure and interaction mechanism of the skin of cashmere goat. At the same time, the data are supplemented for cashmere goat genome resources.
Key words:  Cashmere goat  SMRT  lncRNA  alternative splicing