基于全长转录组测序的牛脂肪组织可变剪接事件分析

Identification of alternative splicing events in bovine adipose tissue based on full-length transcriptome sequencing

为探究可变剪接影响下更为复杂的脂肪沉积调控网络,明晰可变剪接事件的发生对脂肪沉积调控机制的影响,本研究基于PacBio测序平台的第三代测序技术,对夷陵黄牛腹腔脂肪、皮下脂肪、肌间脂肪进行全长转录组测序分析。共发现15445个基因检测到50520个可变剪接,占牛全部基因的33.4%。对这些基因进行富集分析发现,83个GO条目显著富集,这些显著富集的通路可能参与脂肪沉积网络的调控,其中16个与脂质合成及代谢相关,27条KEGG通路显著富集,其中15条与脂质合成及代谢相关。使用KOG、KEGG、NR、Swiss Prot、GO数据库对测序序列进行注释,80756条序列共注释到69259个基因,94458条CDS序列及对应的pep序列。其中GO分析中15039条序列富集到507个生物学过程条目,7907条序列富集到214个细胞组分条目,30672条序列富集到559个分子功能条目。KEGG数据分析富集到34条通路共49710条序列,其中7002条序列参与信号转导途径。以上结果表明,牛脂肪组织存在大量可变剪接事件,并且这些发生可变剪接事件的基因对脂肪沉积的调控发挥重要作用,这可从基因可变剪接角度为解析脂肪沉积的调控网络提供必要的资源和理论依据。

In order to investigate the more complex regulatory network of fat deposition under the influence of alternative splicing,and to clarify the effect of the occurrence of alternative splicing events on the regulatory mechanism of fat deposition,in this study,full-length transcriptome sequencing was performed on abdominal fat,subcutaneous fat and intermuscular fat of Yiling yellow cattle based on the third-generation sequencing technology of PacBio sequencing platform.A total of 50520 alternative splicing were detected in 15445 genes,accounting for 33.4%of all bovine genes.Enrichment analysis of these genes revealed that 83 GO items were significantly enriched,among which 16 were associated with lipid synthesis and metabolism,and 27 KEGG pathways were significantly enriched,among which 15 were associated with lipid synthesis and metabolism.The sequenced sequences were annotated using KOG,KEGG,NR,Swiss Prot,and GO databases,and a total of 80756 sequences were annotated to 69259 genes,94458 CDS sequences and corresponding pep sequences.In GO analysis,15039 sequences were enriched to 507 biological process items,7907 sequences were enriched to 214 cellular component items,and 30672 sequences were enriched to 559 molecular function items.A total of 49710 sequences were enriched in 34 pathways by KEGG data analysis,of which 7002 sequences were involved in signal transduction pathways.The above results indicate that there are a lot of alternative splicing events in bovine adipose tissue,and these genes with alternative splicing events play an important role in the regulation of fat deposition,which may provide necessary resources and theoretical basis for analyzing the regulatory network of fat deposition from the perspective of gene alternative splicing.