Lan DaoLiang, Xiong XianRong, Wei YanLi, Xu Tong, Zhong JinCheng, Zhi XiangDong, Wang Yong, Li Jian
Institute of Qinghai-Tibetan Plateau, Southwest University for Nationalities, Chengdu, 610041, China.
Sci China Life Sci. 2014 Sep;57(9):925-35. doi: 10.1007/s11427-014-4678-2. Epub 2014 Jun 7.
RNA-Seq, a high-throughput (HT) sequencing technique, has been used effectively in large-scale transcriptomic studies, and is particularly useful for improving gene structure information and mining of new genes. In this study, RNA-Seq HT technology was employed to analyze the transcriptome of yak ovary. After Illumina-Solexa deep sequencing, 26826516 clean reads with a total of 4828772880 bp were obtained from the ovary library. Alignment analysis showed that 16992 yak genes mapped to the yak genome and 3734 of these genes were involved in alternative splicing. Gene structure refinement analysis showed that 7340 genes that were annotated in the yak genome could be extended at the 5' or 3' ends based on the alignments been the transcripts and the genome sequence. Novel transcript prediction analysis identified 6321 new transcripts with lengths ranging from 180 to 14884 bp, and 2267 of them were predicted to code proteins. BLAST analysis of the new transcripts showed that 1200?4933 mapped to the non-redundant (nr), nucleotide (nt) and/or SwissProt sequence databases. Comparative statistical analysis of the new mapped transcripts showed that the majority of them were similar to genes in Bos taurus (41.4%), Bos grunniens mutus (33.0%), Ovis aries (6.3%), Homo sapiens (2.8%), Mus musculus (1.6%) and other species. Functional analysis showed that these expressed genes were involved in various Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes pathways. GO analysis of the new transcripts found that the largest proportion of them was associated with reproduction. The results of this study will provide a basis for describing the normal transcriptome map of yak ovary and for future studies on yak breeding performance. Moreover, the results confirmed that RNA-Seq HT technology is highly advantageous in improving gene structure information and mining of new genes, as well as in providing valuable data to expand the yak genome information.
RNA测序(RNA-Seq)是一种高通量测序技术,已有效地应用于大规模转录组学研究,尤其有助于完善基因结构信息和挖掘新基因。在本研究中,采用RNA-Seq高通量技术分析牦牛卵巢转录组。经Illumina-Solexa深度测序后,从卵巢文库中获得了26826516条干净 reads,总长4828772880 bp。比对分析表明,16992个牦牛基因映射到牦牛基因组,其中3734个基因参与可变剪接。基因结构优化分析表明,基于转录本与基因组序列的比对,牦牛基因组中注释的7340个基因在5'或3'端可得到延伸。新转录本预测分析鉴定出6321个长度在180至14884 bp之间的新转录本,其中2267个被预测可编码蛋白质。对新转录本的BLAST分析表明,1200至4933个转录本映射到非冗余(nr)、核苷酸(nt)和/或SwissProt序列数据库。对新映射转录本的比较统计分析表明,它们中的大多数与牛(41.4%)、藏羚羊(33.0%)、绵羊(6.3%)、人(Homo sapiens,2.8%)、小鼠(Mus musculus,1.6%)和其他物种中的基因相似。功能分析表明,这些表达的基因涉及各种基因本体(GO)类别和京都基因与基因组百科全书途径。对新转录本的GO分析发现,其中最大比例与生殖相关。本研究结果将为描绘牦牛卵巢正常转录组图谱及未来牦牛繁殖性能研究提供依据。此外,结果证实RNA-Seq高通量技术在完善基因结构信息、挖掘新基因以及提供有价值的数据以扩展牦牛基因组信息方面具有高度优势。