基于转录组和蛋白质组分析筛选银黑狐毛色形成的关键基因

doi:10.11843/j.issn.0366-6964.2025.09.021

摘要/Abstract

摘要： 旨在筛选影响银黑狐毛色形成的关键基因。在相同饲养条件下，本研究选取9月龄、健康且体重相近的雄性银黑狐和北美赤狐各4只，将其分为银黑狐组(YS)和北美赤狐组(CS)。分别采集两组狐的背中部皮肤样本进行转录组测序(RNA-Seq)和串联质量标签(TMT)标记的定量蛋白质组测序分析，筛选差异表达基因(DEGs)和差异表达蛋白(DEPs)并进行功能富集分析，鉴定与银黑狐毛色形成相关的候选基因。结果表明，与北美赤狐相比，在银黑狐皮肤中共鉴定到2 403个差异表达基因(DEGs)和203种差异表达蛋白质(DEPs)。功能富集分析显示，15个DEGs和6个DEPs通过黑色素合成、PI3K-Akt、cAMP、Wnt和MAPK信号通路参与调控黑色素生物合成。转录组与蛋白质组关联分析表明，TYRP1和NRAS等12个基因在mRNA和蛋白质表达水平上呈现一致的显著变化。蛋白互作网络分析进一步确认了TYRP1、NRAS、MLANA、LEF1、TYR、KITLG、EDNRB、GSK3β、AKT2和ERBB3基因为黑色素合成的关键基因。RT-qPCR和Western blot验证了测序结果的可靠性。本研究确定TYRP1和NRAS可能是影响银黑狐毛色表型的关键候选基因，为银黑狐毛色形成的分子调控机制及狐的分子育种提供了理论依据。

关键词: 狐, 毛色, 转录组, 蛋白质组, 关键基因

Abstract: This study aimed to screen for key genes influencing coat color formation in silver foxes. Under the same feeding conditions, 4 healthy male silver foxes and 4 healthy male red foxes of similar weight at 9 months of age were selected and divided into the silver fox group (YS) and the red fox group (CS). The mid-dorsal skin samples of the two groups were collected for transcriptome sequencing (RNA-Seq) and quantitative proteome sequencing with tandem mass tagging(TMT), and differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were screened and analysed for functional enrichment, to identify candidate genes related to the formation of coat colour in silver foxes. The results showed that a total of 2 403 differentially expressed genes (DEGs) and 203 differentially expressed proteins (DEPs) were identified in the skin of the silver fox compared with the red fox. Functional enrichment analysis revealed that 15 DEGs and 6 DEPs were involved in melanin biosynthesis and metabolic processes through melanin synthesis, PI3K-Akt, cAMP, Wnt, and MAPK signaling pathways. Correlation analysis between transcriptomic and proteomic data indicated that 12 genes, including TYRP1 and NRAS, exhibited consistent significant changes at both mRNA and protein expression levels. Protein-protein interaction (PPI) network analysis further confirmed that TYRP1, NRAS, MLANA, LEF1, TYR, KITLG, EDNRB, GSK3β, AKT2 and ERBB3 were key genes in melanin synthesis. Validation of the sequencing results was performed using RT-qPCR and Western blot analyses. In this study, TYRP1 and NRAS were identified as possible key candidate genes affecting the coat color phenotype of the silver fox, providing a theoretical basis for the molecular regulatory mechanism of coat color formation in the silver fox and for molecular breeding of foxes.

Key words: fox, coat color, transcriptome, proteome, key genes

中图分类号:

S813.3

李佳鹏, 刘庆, 孙佳钰, 马泽芳, 崔凯. 基于转录组和蛋白质组分析筛选银黑狐毛色形成的关键基因[J]. 畜牧兽医学报, 2025, 56(9): 4379-4392.

LI Jiapeng, LIU Qing, SUN Jiayu, MA Zefang, CUI Kai. Screening of Key Genes for Coat Color Formation in Silver Fox Based on Transcriptome and Proteome Analyses[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4379-4392.

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