巴马香猪FGF2基因克隆、生物信息学分析和组织表达研究

doi:10.16174/j.issn.1673-4645.2026.05.008

摘要/Abstract

摘要： 探究成纤维细胞生长因子2（FGF2）在巴马香猪乳腺发育中的生物学功能，明确其序列特征、编码蛋白特性及组织表达规律，为后续功能验证提供基础数据。参考NCBI家猪FGF2基因参考序列（登录号：NM_001405514.1）设计特异性引物，以巴马香猪乳腺组织cDNA为模板，通过RT-PCR克隆FGF2基因的CDS区序列；利用生物信息学软件开展序列比对、进化树构建及编码蛋白结构功能分析；采用qPCR技术检测该基因在巴马香猪不同组织中的表达特征。成功克隆获得巴马香猪FGF2基因CDS区序列，全长468bp，编码155个氨基酸；该序列与家猪同源性达98.7%，在哺乳动物中高度保守，进化上与家猪亲缘关系最近；其编码蛋白为17.25kDa的碱性稳定亲水性蛋白，无信号肽与跨膜结构，存在多个磷酸化修饰位点，可与FGFR家族及乳腺发育关键调控因子互作；FGF2基因在检测组织中呈泛表达特征，乳腺和卵巢中表达量显著高于其他组织（P＜0.05），肝脏中表达量最低。本研究明确了巴马香猪FGF2基因的序列进化、蛋白结构与组织表达规律，为深入探究其在乳腺发育中的分子调控机制提供了理论与试验依据。

关键词: 巴马香猪, FGF2基因, 基因克隆, 生物信息学分析, 乳腺发育

Abstract: This study aimed to explore the biological function of fibroblast growth factor 2 (FGF2) in the mammary gland development of Bama Xiang pigs, clarify its sequence characteristics, encoded protein properties and tissue expression pattern, so as to provide basic data for subsequent functional verification. Specific primers were designed with reference to the reference sequence of porcine FGF2 gene (GenBank accession No.: NM_001405514.1) from NCBI database. The CDS region of FGF2 gene was cloned by RT-PCR using cDNA from mammary gland tissue of Bama Xiang pigs as the template. Sequence alignment, phylogenetic tree construction, and structural and functional analysis of the encoded protein were performed using bioinformatics software. The expression characteristics of FGF2 gene in different tissues of Bama Xiang pigs were detected by qPCR. The CDS sequence of FGF2 gene from Bama Xiang pigs was successfully cloned, with a full length of 468 bp, encoding 155 amino acids. The sequence shared 98.7% homology with that of domestic pigs, was highly conserved in mammals, and had the closest genetic relationship with domestic pigs in evolution. The encoded protein was an alkaline, stable and hydrophilic protein with a molecular weight of 17.25 kDa, without signal peptide and transmembrane domain. It had multiple potential phosphorylation modification sites, and could interact with FGFR family proteins and key regulatory factors of mammary gland development. FGF2 gene showed a ubiquitous expression pattern in the detected tissues, with significantly higher expression levels in the mammary gland and ovary than those in other tissues (P＜0.05), and the lowest expression level in the liver. This study clarified the sequence evolution, protein structure and tissue expression pattern of FGF2 gene in Bama Xiang pigs, which provided a theoretical and experimental basis for further exploring its molecular regulatory mechanism in mammary gland development.

Key words: Bama Xiang pig, FGF2, gene cloning, bioinformatics analysis, mammary gland development

中图分类号: S828.8；S813.3

刘聪, 付张娜, 和艺云, 吴潇潇, 梁爽, 张艳萍, 梁晶. 巴马香猪FGF2基因克隆、生物信息学分析和组织表达研究[J]. 中国猪业, 2026, 21(5): 23-32.

LIU Cong, FU Zhangna, HE Yiyun, WU Xiaoxiao, LIANG Shuang, ZHANG Yanping, LIANG Jing. Cloning, bioinformatics analysis and tissue expression study of FGF2 gene in Guangxi Bama mini-pigs[J]. China Swine Industry, 2026, 21(5): 23-32.

参考文献

[1] 马唯刚, 董钰格, 张长帅, 等. 基于巴马小型猪的智能针灸机器人仿人针刺初步实验[J]. 中国针灸, 2024, 44(12): 1472-1478. MA W G, DONG Y G, ZHANG C S, et al. Preliminary experiment on simulated human manual acupuncture of intelligent acupuncture robot based on Bama miniature pigs[J]. Chinese Acupuncture & Moxibustion, 2024, 44(12): 1472-1478. [2] VICKERS R, PORTER W. Immune cell contribution to mammary gland development[J]. Journal of Mammary Gland Biology and Neoplasia, 2024, 29(1): 16. [3] HANNAN F M, ELAJNAF T, VANDENBERG L N, et al. Hormonal regulation of mammary gland development and lactation[J]. Nature Reviews Endocrinology, 2023, 19(1): 46-61. [4] LIVINGSTON M J, SHU S Q, FAN Y, et al. Tubular cells produce FGF2 via autophagy after acute kidney injury leading to fibroblast activation and renal fibrosis[J]. Autophagy, 2023, 19(1): 256-277. [5] JEONG W, BAE H, LIM W, et al. The functional effects and mechanisms by which fibroblast growth factor 2 (FGF2) controls bovine mammary epithelial cells: implications for the development and functionality of the bovine mammary gland[J]. Journal of Animal Science, 2017, 95(12): 5365-5377. [6] SUMBAL J, JOURNOT R P, ATTRI K, et al. Contractile fibroblasts form a transient niche for the branching mammary epithelium[J]. Nature Communications, 2025, 16: 8576. [7] ZHAO Y, LI M N, HAO Z Y, et al. Effects of FGF2 on characteristics of ovine mammary epithelial cells and milk production traits in dairy sheep[J]. Frontiers in Veterinary Science, 2025, 12: 1709582. [8] SAUCEDO L, SOBARZO C, BRUKMAN N G, et al. Involvement of fibroblast growth factor 2 (FGF2) and its receptors in the regulation of mouse sperm physiology[J]. Reproduction, 2018, 156(2): 163-172. [9] MOR A, MONDAL S, REDDY I J, et al. Molecular cloning and expression of FGF2 gene in pre-implantation developmental stages of in vitro-produced sheep embryos[J]. Reproduction in Domestic Animals, 2018, 53(4): 895-903. [10] 吴双虎, 王鹏飞, 李宏, 等. 关中奶山羊FGF2基因克隆、生物信息学分析及组织表达研究[J]. 中国畜牧兽医, 2022, 49(12): 4554-4563. WU S H, WANG P F, LI H, et al. Cloning, bioinformatics analysis and tissue expression study of FGF2 gene in Guanzhong dairy goats[J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(12): 4554-4563. [11] SUMBAL J, CHICHE A, CHARIFOU E, et al. Primary mammary organoid model of lactation and involution[J]. Frontiers in Cell and Developmental Biology, 2020, 8: 68. [12] DZIEGELEWSKA-SOKOLOWSKA Z, MAHEWSKA A, PROSTEK A, et al. Adipocyte-derived paracrine factors regulate the in vitro development of bovine mammary epithelial cells[J]. International Journal of Molecular Sciences, 2023, 24(17): 13348. [13] 邓丽君, 董晓雪, 牟春燕. FGF/FGFR信号通路调控畜禽毛囊发育与周期性循环的研究进展[J]. 中国畜牧杂志, 2026, 62(3): 101-108. DENG L J, DONG X X, MOU C Y. Research progress on the regulation of hair follicle development and periodic cycle in livestock and poultry by FGF/FGFR signaling pathway[J]. Chinese Journal of Animal Science, 2026, 62(3): 101-108. [14] 王云, 郑彦梓, 唐振华, 等. 广西黄牛FGF2基因克隆、生物信息学分析及其在卵泡中的表达研究[J]. 中国畜牧兽医, 2025, 52(11): 5276-5286. WANG Y, ZHENG Y Z, TANG Z H, et al. Cloning and bioinformatics analysis of FGF2 gene in Guangxi cattle and its expression pattern in ovarian follicles[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5276-5286. [15] 薄双玲, 孙亚洁, 赵欣晨, 等. 成纤维细胞生长因子2对胎鼠肾小体发育的影响[J]. 山西医科大学学报, 2024, 55(8): 970-975. BO S L, SUN Y J, ZHAO X C, et al. Effect of fibroblast growth factor 2 on development of renal corpuscles of embryonic mice[J]. Journal of Shanxi Medical University, 2024, 55(8): 970-975. [16] ZHOU M, SUTLIFF R L, PAUL R J, et al. Fibroblast growth factor 2 control of vascular tone[J]. Nature Medicine, 1998, 4(2): 201-207. [17] 张宇, 黄建芳, 徐萌, 等. FGF2人源化单抗E12及其联合顺铂/紫杉醇体外抑制乳腺癌细胞增殖作用的研究[J]. 中国免疫学杂志, 2018, 34(8): 1187-1194. ZHANG Y, HUANG J F, XU M, et al. Inhibitory effect of novel FGF2 humanized monoclonal antibody E12 combined with cisplatin/paclitaxel on breast cancer cell proliferation in vitro[J]. Chinese Journal of Immunology, 2018, 34(8): 1187-1194. [18] 崔英俊, 于广璞, 汤云飞, 等. 奶牛乳腺分支模型的建立[J]. 中国乳品工业, 2019, 47(1): 16-18. CUI Y J, YU G P, TANG Y F, et al. Establishment of the mammary branching model of dairy cow[J]. China Dairy Industry, 2019, 47(1): 16-18. [19] ZHANG H G, WANG Z F, LIU Z Q, et al. Nuclear FGF2 orchestrates phase separation-mediated rDNA chromatin architecture to control BMSCs cell fate[J]. Bone Research, 2025, 13: 80. [20] KAUR M, LOLICATO F, NICKEL W. Plasma membrane transbilayer asymmetry of PI(4, 5)P2 drives unconventional secretion of Fibroblast Growth Factor 2[J]. Nature Communications, 2025, 16: 10816. [21] XU X X, WANG J Z, GUO X Y, et al. GPR30-mediated non-classic estrogen pathway in mast cells participates in endometriosis pain via the production of FGF2[J]. Frontiers in Immunology, 2023, 14: 1106771. [22] LEI J H, LEE M H, MIAO K, et al. Activation of FGFR2 signaling suppresses BRCA1 and drives triple-negative mammary tumorigenesis that is sensitive to immunotherapy[J]. Advanced Science, 2023, 10(36): e2307935. [23] 刘宣均, 李一兆, 刘开元, 等. IGF-1在哺乳动物卵泡发育中的作用[J]. 畜牧兽医学报, 2026, 57(3): 1177-1186. LIU X J, LI Y Z, LIU K Y, et al. The role of IGF-1 in mammalian follicular development[J]. Acta Veterinaria et Zootechnica Sinica, 2026, 57(3): 1177-1186. [24] YE M H, LI N J, CAI H, et al. Blood and lymphatic vascular network in bone injury repair: from molecular mechanisms to therapeutic strategies[J]. Science China Life Sciences, 2026, 69(7): 2213-2229. [25] 韩俊成, 宋方明, 刘明杰, 等. 基于网络药理学和试验探究金银花治疗奶牛乳房炎的作用机制[J]. 中国兽医学报, 2026, 46(1): 144-155, 174. HAN J C, SONG F M, LIU M J, et al. Investigation of the mechanisms underlying honeysuckle in the treatment of bovine mastitis using network pharmacology and experimental validation[J]. Chinese Journal of Veterinary Science, 2026, 46(1): 144-155, 174. [26] SPERONI L, VOUTILAINEN M, MIKKOLA M L, et al. New insights into fetal mammary gland morphogenesis: differential effects of natural and environmental estrogens[J]. Scientific Reports, 2017, 7: 40806. [27] 崔英俊, 于广璞, 汤云飞, 等. FGF2通过MAPK信号通路影响奶牛乳腺分支形态变化[J]. 东北农业大学学报, 2018, 49(8): 48-53, 72. CUI Y J, YU G P, TANG Y F, et al. FGF2 affects dairy cow mammary epithelial branching morphogenesis through MAPK signaling[J]. Journal of Northeast Agricultural University, 2018, 49(8): 48-53, 72. [28] 王沛婕. PTX3调控奶山羊卵巢颗粒细胞功能及分子机理的研究[D]. 杨凌: 西北农林科技大学, 2021. WANG P J. PTX3 regulates function and molecularmechanism of ovarian granulose cells in dairygoat[D]. Yangling: Northwest A&F University, 2021.

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