牛TGFB1基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2025.06.005

摘要/Abstract

摘要： 【目的】克隆牛转化生长因子β1(transforming growth factor beta 1,TGFB1)基因CDS区，并探究其在固原黄牛不同组织和脂肪细胞分化过程中的表达水平，为后续研究TGFB1基因功能奠定基础。【方法】以牛皮下脂肪组织cDNA为模板，用PCR扩增克隆牛TGFB1基因CDS区。通过生物信息学在线软件预测TGFB1蛋白理化性质、亚细胞定位、蛋白质结构与功能结构域等；利用实时荧光定量PCR检测TGFB1基因在牛脂肪细胞分化中及不同组织中的表达量。【结论】试验成功克隆牛TGFB1基因CDS区，全长1 173 bp。TGFB1基因编码蛋白主要由390个氨基酸组成，蛋白化学分子式为C₁₉₈₁H₃₁₃₄N₅₅₄O₅₆₄S₂₁，总平均亲水性(GRAVY)值为―0.328。TGFB1蛋白共存在37个磷酸化位点、49个O-糖基化位点和3个N-糖基化位点，二级结构主要由α-螺旋和无规则卷曲组成，与SMAD2、SMAD3、ITGAV等蛋白存在互作。实时荧光定量PCR结果显示，与诱导分化0 d相比，在脂肪细胞分化2 d时TGFB1基因表达量显著降低(P＜0.05)，4～10 d时TGFB1基因表达量显著升高(P＜0.05)。组织表达谱显示，TGFB1基因分别在犊牛的脾脏中和成年牛的皮下脂肪中表达量最高，且显著高于其他组织(P＜0.05)。【结论】牛TGFB1基因在牛前体脂肪细胞诱导分化4 d时表达量较高，且在犊牛脾脏和成年牛皮下脂肪中高表达，其在调控脂肪细胞增殖和分化过程及犊牛免疫系统的生理过程中发挥重要作用。研究结果为进一步探究TGFB1基因参与固原黄牛脂肪沉积的分子机制提供了理论基础。

关键词: 固原黄牛; TGFB1基因; 牛前体脂肪细胞; 组织表达

Abstract: 【Objective】 The aim of this experiment was to clone the CDS region of bovine transforming growth factor beta 1 (TGFB1) gene and explore its expression in different tissues and adipocytes during differentiation of Guyuan cattle,laying a foundation for the subsequent research on the function of TGFB1 gene.【Method】 The CDS region of bovine TGFB1 gene was amplified by PCR using adipose tissue cDNA as template.The physicochemical properties,subcellular localization,protein structure and functional domain of TGFB1 protein were predicted by bioinformatics online softwares.Real-time quantitative PCR was used to detect the expression of TGFB1 gene in bovine adipocyte during differentiation and different tissues.【Result】 The CDS region of bovine TGFB1 gene was successfully cloned with a total length of 1 173 bp.The protein encoded by TGFB1 gene was mainly composed of 390 amino acids and its chemical formula was C₁₉₈₁H₃₁₃₄N₅₅₄O₅₆₄S₂₁.The total average hydrophilic (GRAVY) value was ―0.328.TGFB1 protein had 37 phosphorylation sites,49 O-glycosylation sites and 3 N-glycosylation sites.The secondary structure of TGFB1 protein was mainly composed of alpha helix and random coil,and it interacted with SMAD2,SMAD3,ITGAV and other proteins.Real-time quantitative PCR results showed that compared with 0 d after induction differentiation，TGFB1 gene expression was significantly decreased at 2 d (P＜0.05),and significantly increased at 4-10 d (P＜0.05).Tissue expression profile showed that the expression of TGFB1 gene was the highest in spleen of calves and subcutaneous fat of adult cows,respectively,and was significantly higher than that in other tissues (P＜0.05). 【Conclusion】 The expression level of the bovine TGFB1 gene was relatively high at 4 d of bovine precursor adipocytes, and it was highly expressed in spleen of calves and subcutaneous fat of adult calves. It played an important role in regulating the proliferation and differentiation of adipocytes and the physiological processes of the calf immune system. This results provided a theoretical basis for further exploring the molecular mechanism of the TGFB1 gene involved in fat deposition in Guyuan cattle.

Key words: Guyuan cattle; TGFB1 gene; bovine preadipocytes; tissue expression

中图分类号:

S823

楚洪恩, 刘源, 冯雪, 白雪, 杨梦丽, 李娟, 贺丽霞, 刘爽, 冯兰, 马云. 牛TGFB1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(6): 2506-2518.

CHU Hongen, LIU Yuan, FENG Xue, BAI Xue, YANG Mengli, LI Juan, HE Lixia, LIU Shuang, FENG Lan, MA Yun. Cloning,Bioinformatics and Tissue Expression Analysis of Bovine TGFB1 Gene[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2506-2518.

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