| 本文已被：浏览 396次 下载 59次 |
码上扫一扫！ |
| 牦牛FGF6基因克隆及组织表达特异性研究 |
|
刘毅程1,2, 杨焱3, 洛桑顿珠4, 马晓明1,4, 卓玛次仁5, 尼玛加措5, 平措占堆4, 梁春年1,4
|
|
1.中国农业科学院 兰州畜牧与兽药研究所，甘肃省牦牛繁育工程重点实验室，农业农村部青藏高原畜禽遗传育种重点实验室，甘肃 兰州 730050;2.甘肃农业大学 动物科学技术学院，甘肃 兰州 730050;3.肃南县农业农村局，甘肃 肃南 734400;4.西藏自治区农牧科学 院 畜牧兽医研究所，西藏 拉萨 850004;5.日喀则市昂仁县农业农村局，西藏 日喀则 857001
|
|
| 摘要: |
| 【 目的】克隆牦牛成纤维细胞生长因子6（FGF6）基因，对其进行生物信息学分析及组织表达分析，初 步揭示该基因的结构和功能，为FGF6基因在牦牛中的功能研究提供理论依据。【 方法】选取3头桑桑牦牛公牛，屠宰 后采集脂肪、心脏、肝脏、肌肉、肺脏、肾脏及脾脏组织，提取脂肪组织总RNA，反转录获得cDNA。利用RT-PCR技术克隆牦牛FGF6基因的CDS区序列，使用多种软件对该基因进行生物信息学分析。使用RT-qPCR技术检测FGF6基因在桑桑牦牛脂肪、心脏、肝脏、肌肉、肺脏、肾脏及脾脏组织中的表达情况。【 结果】牦牛FGF6基因的CDS区全长为627 bp，共编码208个氨基酸。牦牛FGF6蛋白分子式为C1010H1619N291O285S7，分子质量为22 623. 16 u，理论等电点为10. 24，脂肪系数（AI）为89.04，总平均亲水指数（GRAVY）为?0.083，不稳定指数为36.86，表明该蛋白为稳定型亲水 性蛋白；FGF6蛋白在第44~45位氨基酸处存在一个信号肽，在氨基酸序列的20~42位处存在一个跨膜螺旋；该蛋白 存在1个N-糖基化位点和22个磷酸化位点。亚细胞定位结果显示，牦牛FGF6蛋白分布于线粒体、细胞质、高尔基体、 内质网、囊泡、细胞外，其氨基酸组成中甘氨酸含量最高（12%），蛋白二级结构主要以无规则卷曲为主（占64.9%）。同 源性比对发现，桑桑牦牛与野牦牛的亲缘关系最近（100%），其次为瘤牛（99.9%），与马鹿的亲缘关系最远（72.8%）；FGF6基因与FGFR1、FGFR2、FGFR4等相关蛋白存在相互作用。RT-qPCR结果显示，FGF6基因在桑桑牦牛不同 组织中均有表达，其中在肌肉组织中表达量最高，极显著高于其他组织（P<0.01），在心脏的表达量极显著高于脂肪、 肝脏、肺脏、肾脏、脾脏（P<0.01）。【 结论】获得了牦牛FGF6基因完整的CDS区，发现其在不同物种间保守性较高。 |
| 关键词: 牦牛 FGF6基因 生物信息学分析 |
| DOI：10. 13207/j. jnwafu. 2026. 08. 001 |
| 分类号: |
| 基金项目:中央引导地方科技发展资金项目（25ZYJA0）；西藏自治区重点研发计划项目（XZ202402ZY0012）；现代肉牛牦牛产业技术体 系项目（CARS-37） |
|
| Cloning of yak FGF6 gene and study on its tissue expression specificity |
|
LIU Yicheng1,2, YANG Yan3, Lobsang Dunzhu4, MA Xiaoming1,4, ZhuomaTsering5, Nima Gyatso5, Phuntsok Zhandui4, LIANG Chunnian1,4
|
|
1.Lanzhou Institute of Husbandry and Pharmaceutical Sciences，Gansu Provincial Key Laboratory of Yak Breeding Engineering，Key Laboratory of Livestock and Poultry Genetics and Breeding on Qinghai-Tibet Plateau，Ministry of Agriculture and Rural Affairs，Lanzhou Institute of Husbandry and Pharmaceutical Sciences，Chinese Academy of Agricultural Sciences，Lanzhou，Gansu 730050，China;2.College of Animal Science and Technology，Gansu Agricultural University，Lanzhou，Gansu 730050，China;3.Agriculture and Rural Affairs Bureau of Sunan County，Sunan，Gansu 734400，China;4.Institute of Animal Husbandry and Veterinary Medicine，Xizang Academy of Agricultural and Animal Husbandry Sciences，Lhasa，Xizang 850004，China;5.Agriculture and Rural Affairs Bureau of Angren County，Shigatse，Xizang 857001，China
|
| Abstract: |
| 【 Objective】This research aims to clone the yak fibroblast growth factor 6（FGF6） gene，con?duct bioinformatic analysis and tissue expression analysis，and initially reveal the structure and function of the gene，so as to provide a theoretical basis for the functional study of FGF6 gene in yak.【Method】Three Sang?sang yak bulls were selected，and their adipose，heart，liver，muscle，lung，kidney and spleen tissues were collected after slaughter. Total RNA of adipose tissue was extracted，and the cDNA was obtained through reverse transcription. The sequence of the CDS region of the FGF6 gene of yaks was cloned by using RT-PCR，and multiple tools were employed to conduct bioinformatic analysis on the FGF6 gene. RT-qPCR was used to detect the expression level of the FGF6 gene in adipose，heart，liver，muscle，lung，kidney and spleen tissue of Sangsang yaks.【Result】The CDS region of the yak FGF6 gene was 627 bp in length，encoding a total of 208 amino acids. The yak FGF6 protein molecular composition was C1010H1619N291O285S7，molecular mass was 22 623.16 u，theoretical isoelectric point was 10.24，aliphaticity coefficient（AI） was 89.04，total average hydrophilicity index（GRAVY） was -0.083，and instability index was 36.86，which indicated that the protein was a stable hydrophilic protein. A signal peptide was observed at amino acid position 44-45 in FGF6 protein，and a transmembrane helix at amino acid position 20-42.One N-glycosylation site and 22 phosphorylation sites were identified in the protein. The results of subcellular localization showed that the yak FGF6 protein was distributed in mitochondria，cytoplasm，Golgi，endoplasmic reticulum，vesicles，and extracellular part. Among the amino acids of the encoded protein，glycine showed the highest proportion （12%），and the secondary structure of the pro?tein was predominantly irregular coiling （64.9%）. Homology comparison revealed that the Sangsang yak was most closely related to the wild yak（100%），followed by the Bos indicus （99.9%） and least closely related to the horse deer（72.8%）.Interaction was observed between the FGF6 gene and its related proteins，such as FGFR1，FGFR2，FGFR4，etc. The results of RT-qPCR showed that the FGF6 gene was expressed in different tissues of the Sangsang yak，with the highest expression level observed in the muscle tissue，which was highly significantly higher than those in other tissues（P<0. 01）. And the expression level in heart tissue was highly sig?nificantly higher than those in adipose，liver，lung，kidney and spleen tissues（P<0. 01）.【Conclusion】The com?plete CDS region of yak FGF6 gene was obtained and found to be highly conserved among different species. |
| Key words: yak FGF6 gene bioinformatics analysis |
