山羊INSR基因可变剪接体在背最长肌中的表达模式

doi:10.11843/j.issn.0366-6964.2020.08.006

摘要/Abstract

摘要： 旨在对山羊胰岛素受体（insulin receptor，INSR）基因进行可变剪接分析，研究其在背最长肌中的表达模式。本试验以简州大耳羊妊娠期胎儿（妊娠期第45、60和105天）及出生后第3天羔羊背最长肌为试验材料，基于转录组测序数据（accession number：SRR3567144）并利用Sanger测序验证，分析INSR基因的可变剪接体类型及其在背最长肌中的表达差异，同时利用在线软件对不同可变剪接体进行生物信息学分析。结果显示，在山羊背最长肌中INSR基因CDS区存在4种不同的可变剪接形式（INSR1、INSR2、INSR3和INSR4），其CDS长分别为4 110、4 146、4 113和4 149 bp，对应的氨基酸残基数分别为1 369、1 381、1 370和1 382个。山羊INSR 4个可变剪接体之间的主要区别在于Exon 11（36 bp）跳跃和Exon 15部分（3 bp）缺失，这两种可变剪接模式在牛、猪、山羊、人、绵羊和小鼠间完全一致。这些可变剪接改变了山羊INSR的Ser磷酸化位点和第二个FN3功能域，使所编码蛋白3D结构在两个区域发生明显改变。同时，各可变剪接体表达量在背最长肌发育的4个时期均无显著性差异（P>0.05），同一时期各可变剪接体表达量之间也无显著性差异（P>0.05）。INSR基因序列和剪接模式在哺乳动物中高度保守，该研究结果将为深入揭示INSR基因对动物肌肉发育的调控机制提供理论参考。

关键词: 山羊, INSR, 可变剪接体, 表达模式

Abstract: The study was conducted to perform alternative splicing analysis on goat insulin receptor (INSR) gene, and to analyze its expression profiles in goat Longissimus dorsi muscle. The Longissimus dorsi muscle of Jianzhou Daer goat were collected from fetuses at 45, 60, and 105 days of gestation and lambs at the 3rd day after birth. Based on Sanger sequencing and high throughput sequencing data (accession number: SRR3567144), the alternative splicing patterns of the INSR gene and their expression difference in Longissimus dorsi muscle were studied. In addition, the characteristics of different alternative splicing variants of goat INSR gene were analyzed using online softwares. The results showed that 4 different INSR transcripts were identified and designated as INSR1, INSR2,INSR3 and INSR4, with CDS lengths of 4 110, 4 146, 4 113 and 4 149 bp, respectively, and the correspondingly amino acid residues were 1 369, 1 381, 1 370 and 1 382. The Exon 11 (36 bp) skipping and partial deletion of Exon 15 (3 bp) led to the main difference among 4 transcripts of INSR in goat. The two alternative splicing patterns were completely identical among INSR gene transcripts of bovine, pig, goat, human, sheep and mouse. These alternative splicing changed the serine phosphorylation sites and the second FN3 domain of goat INSR, and resulted in 3D structure change of the encoded protein in two regions. Meanwhile, there was no significant difference in the expression level of 4 splicing variants during 4 Longissimus dorsi muscle developmental periods of goats (P>0.05), and there was no significant difference among the expression level of 4 splicing variants within the same developmental period (P>0.05). In conclusion, the sequence and splicing model of INSR gene's splicing variants are highly conserved among mammals, and the results have provided the theoretical reference for further revealing the role of INSR gene in regulating animal muscle development.

Key words: goat, INSR, splicing transcripts, expression patterns

中图分类号:

S827.2

张潇, 李利, 张红平. 山羊INSR基因可变剪接体在背最长肌中的表达模式[J]. 畜牧兽医学报, 2020, 51(8): 1823-1832.

ZHANG Xiao, LI Li, ZHANG Hongping. Identification of Splicing Transcripts of Insulin Receptor Gene and Their Expression Profiles in Goat Longissimus Dorsi Muscles[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1823-1832.

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