九疑山兔FGF10基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2025.03.003

摘要/Abstract

摘要： 【目的】探讨九疑山兔成纤维细胞生长因子10(fibroblast growth factor 10，FGF10)基因的分子结构特征及其在九疑山兔不同日龄、不同组织中的表达模式。【方法】试验以九疑山兔背最长肌为材料，对九疑山兔FGF10基因编码区序列(CDS)进行克隆，利用MegAlign软件分析FGF10氨基酸序列相似性，并使用Mega 11.0软件构建系统进化树。利用ProtParam、SOPMA等在线网站对FGF10蛋白的理化性质、二级结构和三级结构等进行预测分析，利用实时荧光定量PCR技术检测FGF10基因在1、35、84、180日龄九疑山兔的心脏、肝脏、脾脏、肺脏、肾脏、背最长肌中的表达量。【结果】九疑山兔FGF10基因CDS区长度为663 bp，共编码220个氨基酸，其编码的氨基酸序列与家兔、欧洲兔、鸡、蟾蜍、绵羊、家牛、马、人、小鼠的相似性分别为100.0%、99.5%、87.3%、73.1%、97.2%、97.2%、99.1%、99.0%、93.8%，系统进化树结果显示九疑山兔与马的亲缘关系较近。九疑山兔FGF10蛋白属于亲水性不稳定蛋白，存在1个跨膜区域和1个信号肽，其结构域包括跨膜区、低复杂度区域和FGF家族同源结构域等结构，并含有2个糖基化位点和40个磷酸化位点。其二级结构以无规则卷曲为主(49.55%)，其次为延伸链(25.91%)，三级结构与二级结构的预测结果一致。蛋白互作预测发现FGF10蛋白还与KL、FGFR2、FGFR4等蛋白存在互作。实时荧光定量PCR结果表明，FGF10基因在4个日龄九疑山兔的肺脏中表达量均为最高，肾脏其次，而在肝脏中表达量最低。此外，FGF10基因在九疑山兔背最长肌组织中的表达水平随着时间的延长呈现下降趋势，在84日龄时达到最低点。【结论】九疑山兔FGF10蛋白与家兔和欧洲兔的相似性最高，为不稳定的亲水蛋白，存在跨膜区、信号肽、糖基化位点和磷酸化位点。FGF10基因在九疑山兔不同日龄、不同组织中差异表达。试验结果为深入研究FGF10基因在肌肉发育中的潜在作用提供了重要的参考依据。

关键词: 九疑山兔; FGF10基因; 克隆; 分子特性; 时空表达谱

Abstract: 【Objective】 The aim of this study was to explore the molecular structural characteristics of fibroblast growth factor 10 (FGF10) gene in Jiuyishan rabbits and its expression levels in different tissues of Jiuyishan rabbits at different ages.【Method】 The longissimus dorsi muscle of Jiuyishan rabbit was used as material in the experiment to clone the coding region sequence (CDS) of FGF10 gene in Jiuyishan rabbits.The amino acid sequence similarity of FGF10 was analyzed with MegAlign software，and a phylogenetic tree was constructed using Mega 11.0 software.The physicochemical properties,secondary structure,and tertiary structure of the FGF10 protein were predicted and analyzed using online tools like ProtParam and SOPMA.The expression of FGF10 gene in heart,liver,spleen,lung,kidney and longissimus dorsi muscle of 1,35,84 and 180 days old Jiuyishan rabbits was detected by Real-time quantitative PCR.【Result】 The length of CDS of FGF10 gene in Jiuyishan rabbits was 663 bp,which encoded 220 amino acids.The similarity of its encoded amino acid sequence with Oryctolagus cuniculus，Lepus europaeus，Gallus domestiaus,Pelobates fuscus，Ovis aries，Bos taurus，Equus caballus，Homo sapiens and Mus musculus by 100.0%,99.5%,87.3%,73.1%,97.2%,97.2%,99.1%,99.0% and 93.8%,respectively.The results of the phylogenetic tree showed that the genetic relationship between Jiuyishan rabbits and Equus caballus was relatively close. FGF10 protein of Jiuyishan rabbits was characterized as an hydrophilic unstable protein.It was characterized by the presence of a transmembrane region and a signal peptide.The structural domains of the FGF10 protein included a transmembrane region,a low-complexity region,and a homologous domain that was typical of the FGF family,in addition to two sites designated for glycosylation and 40 sites for phosphorylation.Its secondary structure was predominantly composed of random coils (49.55%),with extended chains making up another 25.91%.The predicted tertiary structure aligned with the secondary structure.Protein interaction predictions revealed that FGF10 protein interacted with KL,FGFR2,FGFR4,and other proteins.Real-time quantitative PCR results revealed that the expression of FGF10 gene at four stages was the highest in lung,followed by kidneys,and the lowest in liver.In addition,the expression of FGF10 gene in longissimus dorsi muscle showed a downward trend with time,and reached the lowest level at 84 days of age.【Conclusion】 FGF10 protein from Jiuyishan rabbits had the highest similarity with Oryctolagus cuniculus and Lepus europaeus and was an unstable hydrophilic protein with transmembrane regions,signal peptides,glycosylation sites,and phosphorylation sites. FGF10 gene was differentially expressed in different ages and tissues of Jiuyishan rabbits.These findings provided important reference for further studying the potential role of FGF10 gene in muscle development.

Key words: Jiuyishan rabbits; FGF10 gene; cloning; molecular characteristic; spatiotemporal expression spectrum

中图分类号:

S829.1

黄念旎, 黄子珂, 李聪聪, 刘晗, 马诗语, 宋果, 蒋亮, 彭为波, 杨联熙, 黄生强. 九疑山兔FGF10基因克隆、生物信息学及组织表达分析[J]. 中国畜牧兽医, 2025, 52(3): 1001-1010.

HUANG Nianni, HUANG Zike, LI Congcong, LIU Han, MA Shiyu, SONG Guo, JIANG Liang, PENG Weibo, YANG Lianxi, HUANG Shengqiang. Cloning,Bioinformatics and Tissue Expression Analysis of FGF10 Gene in Jiuyishan Rabbits[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1001-1010.

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