关中奶山羊FGF2基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2022.12.005

Abstract

Abstract: 【Objective】 The purpose of this study was to perform cloning and bioinformatic analysis of fibroblast growth factor 2 (FGF2) gene in Guanzhong dairy goats, and detect the expression differences of FGF2 gene in various tissues of goats, so as to lay the foundation for the subsequent exploration of the function of FGF2 gene.【Method】 Using mammary gland cDNA of Guanzhong dairy goat as template, the complete CDS region of FGF2 gene was amplified by RT-PCR and cloned, the similarity alignment, phylogenetic tree and bioinformatics analysis were carried out.Meanwhile, Real-time quantitative PCR was used to detect the expression of FGF2 gene in heart, liver, spleen, kidney, longissimus dorsi muscle, lung, mammary gland and ovary of Guanzhong dairy goats.【Result】 The coding region of FGF2 gene in Guanzhong dairy goats was 468 bp, encoding 155 amino acids, the molecular mass was 17.26 ku and the theoretical isoelectric point was 9.58, the highest content was glycine, accounting for 10.3%.The similarity alignment results showed that the similarity of FGF2 amino acid sequence of Guanzhong dairy goat with Homo sapiens, Bos taurus, Equus caballus, Oryctolagus cuniculus, Ovis aries, Orcinus and Sus scrofa were 98.7%, 99.4%, 100.0%, 98.7%, 99.4%, 99.4% and 62.6%, respectively.The phylogenetic tree showed that FGF2 gene was highly conserved among different species and had the closest homologous relationship with Equus caballus.The instability index of FGF2 protein of Guanzhong dairy goat was 38.39, which was an stable hydrophilic protein without signal peptide and transmembrane domain.The secondary structure of FGF2 protein contained alpha helix, beta turn, extended chain and random coil, accounting for 10.32%, 14.19%, 30.97% and 44.52%, respectively.Protein interaction prediction analysis found that FGF2 protein interacted with FGFR, KDR, FGF1, FGFR4, MET and FGFR1 proteins related to mammary gland growth and development.Real-time quantitative PCR detected that FGF2 gene of Guanzhong dairy goat was expressed in heart, liver, spleen, kidney, longissimus dorsi muscle, lung, mammary gland and ovary.The expression in mammary gland was highest, followed by ovary, and the lowest expression in longissimus dorsi muscle.【Conclusion】 The CDS region of FGF2 gene sequence in Guanzhong dairy goat was 468 bp, and encoded 155 amino acids, which was a hydrophilic protein.The secondary structure of FGF2 protein was mainly extended chain and random coil.FGF2 gene in Guanzhong dairy goat was expressed in different tissues during the peak lactation period.The results of this study provided a reference for further exploring the role of FGF2 gene in Guanzhong dairy goat mammary gland development and its specific functions.

Key words: Guanzhong dairy goats; FGF2 gene; cloning; bioinformatics analysis; tissue expression

CLC Number:

S827

WU Shuanghu, WANG Pengfei, LI Hong, CHAO Juanjuan, WANG Zhanhang, ZHU Junru, AN Xiaopeng. Cloning, Bioinformatics Analysis and Tissue Expression Study of FGF2 Gene in Guanzhong Dairy Goats[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4554-4563.

References

[1] IDRISOVA K F, ZEINALOVA A K, MASGUTOVA G A, et al.Application of neurotrophic and proangiogenic factors as therapy after peripheral nervous system injury[J].Neural Regeneration Research, 2022, 17(6):1240-1247.
[2] SEETH M T, HOEDEMAKER M, KROMKER V.Physiological processes in the mammary gland tissue of dairy cows during the dry period[J].Berliner und Munchener Tierarztliche Wochenschrift, 2015, 128(1-2):76-83.
[3] 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.
[4] 于广璞.FGF2调节奶牛乳腺分支的分子机理研究[D].哈尔滨:东北农业大学, 2018. YU G P.Molecular mechanism of FGF2 regulating mammary gland branching in dairy cows[D].Harbin:Northeast Agricultural University, 2018.(in Chinese)
[5] 董飞.奶山羊乳腺组织差异表达miRNA功能预测及miR-135a对PRLR基因靶向调控研究[D].泰安:山东农业大学, 2014. DONG F.Functional prediction of differentially expressed miRNAs in milk goat mammary gland tissue and the targeted regulation of PRLR gene by miR-135a[D].Tai'an:Shandong Agricultural University, 2014.(in Chinese)
[6] AVAGLIANO A, FIUME G, RUOCCO M R, et al.Influence of fibroblasts on mammary gland development, breast cancer microenvironment remodeling, and cancer cell dissemination[J].Cancers, 2020, 12(6):1697.
[7] SUMBAL J, KOLEDOVA Z.FGF signaling in mammary gland fibroblasts regulates multiple fibroblast functions and mammary epithelial morphogenesis[J].Development, 2019, 146(23):185306.
[8] 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.
[9] 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.
[10] WONG C G, TABAN M, OSANN K, et al.Subchoroidal release of VEGF and bFGF produces choroidal neovascularization in rabbit[J].Current Eye Research, 2017, 42(2):237-243.
[11] HOTA J, MAHAPATRA P K. FGF2 and FGF10 expression patterns in the epidermis and mesenchyme during homeotic transformation of tail into hindlimbs in frog tadpoles[J].Acta Histochemica, 2021, 124(1):151836.
[12] SUDHA T, MOUSA D S, EL-FAR A H, et al.Pomegranate (Punica granatum) fruit extract suppresses cancer progression and tumor angiogenesis of pancreatic and colon cancer in chick chorioallantoic membrane model[J].Nutrition and Cancer-An International Journal, 2021, 73(8):1350-1356.
[13] CHOI K H, LEE D K, OH J N, et al.FGF2 signaling plays an important role in maintaining pluripotent state of pig embryonic germ cells[J].Cellular Reprogramming, 2018, 20(5):301-311.
[14] KUMAR S, SINGLA S K, MANIK R, et al.Effect of basic fibroblast growth factor (FGF2) on cumulus cell expansion, in vitro embryo production and gene expression in buffalo (Bubalus bubalis)[J].Reproductive Biology, 2020, 20(4):501-511.
[15] 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.
[16] 江楠楠, 刘建新, 石恒波.乳腺脂肪组织调节乳腺发育的研究进展[J].生物化学与生物物理进展, 2021, 48(6):611-617. JIANG N N, LIU J X, SHI H B.Research progress on the regulation of mammary gland development by mammary adipose tissue[J].Advances in Biochemistry and Biophysics, 2021, 48(6):611-617.(in Chinese)
[17] QUALMANN B, KESSELS M M.The role of protein arginine methylation as post-translational modification on actin cytoskeletal components in neuronal structure and function[J].Cells, 2021, 10(5):1079.
[18] STERINGER J P, NICKEL W.A direct gateway into the extracellular space:Unconventional secretion of FGF2 through self-sustained plasma membrane pores[J]. Seminars in Cell & Developmental Biology, 2018, 83:3-7.
[19] LABRCHE C, COOK D P, ABOU-HAMAD J, et al.Periostin gene expression in neu-positive breast cancer cells is regulated by a FGFR signaling cross talk with TGFβ/PI3K/Akt pathways[J].Breast Cancer Research:BCR, 2021, 23(1):107.
[20] GAO Y, WANG Y W, YU J, et al.FGF exhibits an important biological role on regulating cell proliferation of breast cancer when it transports into the cell nuclei[J].Cell Biochemistry and Biophysics, 2021, 80(2):311-320.
[21] PARADOWSKA-GORYCKA A, STYPINSKA B, PAWLIK A, et al.KDR (VEGFR2) genetic variants and serum levels in patients with rheumatoid arthritis[J].Biomolecules, 2019, 9(8):355.
[22] MOHAMMAD R F, HASHEMZADEH S, RAVANBAKHSH G R, et al.Dysregulated KDR and FLT1 gene expression in colorectal cancer patients[J].Reports of Biochemistry and Molecular Biology, 2019, 8(3):244-252.
[23] GUO R B, BERRY L D, AISNER D L, et al.MET IHC is a poor screen for MET amplification or MET exon 14 mutations in lung adenocarcinomas:Data from a Tri-institutional cohort of the lung cancer mutation consortium[J].Journal of Thoracic Oncology, 2019, 14(9):1666-1671.
[24] AREF-ESHGHI E, LIN F M, LI M R M, et al.The oncogenic roles of NTRK fusions and methods of molecular diagnosis[J].Cancer Genetics, 2021, 258:110-119.
[25] 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 the function and molecular mechanism of ovarian granulosa cells in dairy goats[D].Yangling:Northwest A&F University, 2021.(in Chinese)
[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. CUI Y J, YU G P, TANG Y F, et al.FGF2 affects the morphological changes of dairy cow mammary gland branches through MAPK signaling pathway[J].Journal of Northeast Agricultural University, 2018, 49(8):48-53.(in Chinese)
[28] POND A C, BIN X, BATTS T, et al.Fibroblast growth factor receptor signaling is essential for normal mammary gland development and stem cell function[J].Stem Cells, 2013, 31(1):178-189.

Cloning, Bioinformatics Analysis and Tissue Expression Study of FGF2 Gene in Guanzhong Dairy Goats

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIU Jialong, ZHANG Yiwen, ZHANG Yueran, SUN Qijuan, CHEN Jian, HE Lei, JIA Yanyan, DING Ke, YU Zuhua. Cloning,Prokaryotic Expression and Bioinformatics Analysis of ELAVL1 Gene in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1807-1817.
[2]	ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.
[3]	TAO Qinghua, HUANG Caiyun, CHEN Chao, LIU Wei, SUN Ningning, LI Ang. Cloning and Tissue Expression Difference Analysis of LIF Gene in White Muscovy Duck [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1296-1306.
[4]	FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.
[5]	LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.
[6]	HU Leyu, HU Xinyao, LI Ying, ZHAO Shengmiao, SHEN Fengzhen, WANG Changfa, LI Liangliang, WANG Tongtong, REN Huiying. Bioinformatics Analysis of HO-1 Gene and Preparation of Polyclonal Antibody in Dezhou Donkey [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1499-1510.
[7]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[8]	LI Dan, ZHANG Haisen, WANG Yiqun, GAO Dengke, ZHAO Hongcong, LI Chao, JIN Yaping, CHEN Huatao. Eukaryotic Expression Vector Construction,Bioinformatics and Tissue Expression Profiles Analysis of CRY1 Gene in Dairy Cows [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 870-881.
[9]	ZHANG Yongjie, SUI Zhiyuan, ZHANG Zhishuai, WANG Chenguang, LI Xiaojun, XING Feng. Cloning and Bioinformatics Analysis of NPTX1 Gene and Its Tissues Expression at Puberty in Dolang Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1048-1058.
[10]	WEI Yirong, ZHENG Zihua, ZHANG Sanbao, SONG Ying, JIANG Haiyu, SUN Wenyue, CHENG Pengfei, LIU Yufan, ZOU Jianwei, HUANG Yanna, PAN Yan, JIANG Qinyang. Cloning,Bioinformatics Analysis and Tissue Expression of UCP1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 440-450.
[11]	WANG Guang, ZOU Jiahao, ZHANG Yongtao, LI Dexian, LI Xueqing, YU Mengqi, CHEN Lu, YUAN Yuxin, LI Guang. Difference Analysis of Metabolites in Testis of Guanzhong Dairy Goats at Different Developmental Stages [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3453-3464.
[12]	TANG Jingyu, DU Hanyu, MENG Chunchun, LIU Guangqing. Cloning,Bioinformatics Analysis and Subcellular Localization of Interferon-stimulated Gene 15 in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2843-2854.
[13]	FU Yongzhi, MA Xiaojun, WANG Maohan, LI Zhe, CHEN Zhiyuan, QUAN Wei, ZHAO Han, SU Yongmei, WU Jianhua, ZHU Wenjin. Bioinformatics Analysis and Tissue Expression of MSTN Gene in Yangyuan Donkeys [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2910-2919.
[14]	QI Nannan, QIAN Yonghang, LUO Gang, SITU Xuqi, LIU Jiying. Cloning, Bioinformatics Analysis and Tissue Expression Location of Porcine MORC2 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2431-2441.
[15]	ZHANG Hang, SHA Huiyang, LI Huawei, HUANG Liangzong, ZHAO Mengmeng. Cloning, Bioinformatics and Tissue Expression Analysis of TRIM3 Gene in Swine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2474-2483.