中国畜牧兽医2025，Vol.52Issue(11)：5276-5286,11.DOI:10.16431/j.cnki.1671-7236.2025.11.024
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
Cloning and Bioinformatics Analysis of FGF2 Gene in Guangxi Cattle and Its Expression Pattern in Ovarian Follicles
摘要
Abstract
[Objective]This study aimed to clone the coding sequence(CDS)of the fibroblast growth factor 2(FGF2)gene in Guangxi cattle and conduct bioinformatics analysis,investigate the expression patterns of FGF2 gene in various tissues and ovarian follicles,and establish a foundation for further exploration of its functional roles in follicular development.[Method]The FGF2 gene fragment was amplified from Guangxi cattle ovarian cDNA,followed by sequence alignment and phylogenetic tree construction with other species.Bioinformatics analysis of FGF2 protein was performed using online tools.The expression of FGF2 gene in different tissues of Guangxi cattle were detected via Real-time quantitative PCR.ELISA was employed to quantify FGF2 concentrations in follicular fluid from small(3-4 mm diameter),medium(5-8 mm),and large(＞8 mm)follicles.Immunofluorescence and Real-time quantitative PCR were combined to analyze the spatial localization and expression dynamics of FGF2 gene in follicular cells.[Result]The CDS of FGF2 gene in Guangxi cattle spanned 468 bp,encoding 155 amino acids.The amino acid sequence similarity of FGF2 protein in Guangxi cattle exhibited＞98％with Sus scrofa,Capra hircus,Ovis aries,and Oryctolagus cuniculus,demonstrating high conservation among mammals.Bioinformatics analysis results revealed that FGF2 was an alkaline hydrophilic protein containing 14 phosphorylation sites,with secondary structures dominated by random coil(61.94％)and extended chain(29.03％),and key interacting partners included FGFR,PDGFRA,GPC1,and EGFR.Tissue expression analysis revealed that FGF2 gene was expressed in various tissues of Guangxi cattle,with significantly higher expression in ovarian and lung tissues compared with other tissues(P＜0.05).Follicular fluid FGF2 concentrations in large follicles(＞8 mm)were significantly elevated relative to small follicles(3-4 mm)(P＜0.05).The expression of FGF2 gene in oocytes was significantly higher than that in granulosa and theca cells(P＜0.05),while its receptor FGFR1 gene expression in oocytes was significantly lower than that in granulosa and theca cells(P＜0.05).[Conclusion]The CDS region of FGF2 gene in Guangxi cattle was 468 bp,encoding a polypeptide of 155 amino acids characterized as a stable alkaline hydrophilic protein,with its secondary structure dominated by random coil.FGF2 gene exhibited significantly higher expression in ovarian and lung.Within ovarian follicles,its expression was predominantly localized to oocytes,and it regulated follicular development via follicle diameter-dependent concentration dynamics.The results provided a theoretical foundation for elucidating the functional regulatory network of FGF2 gene in ovaries of Guangxi cattle.关键词
广西黄牛/FGF2基因/克隆/生物信息学/表达模式/卵泡发育Key words
Guangxi cattle/FGF2 gene/cloning/bioinformatics/expression pattern/follicle development分类
农业科技引用本文复制引用
王云,石德顺,陆凤花,郑彦梓,唐振华,王燕新,贾茹茹,韦春烨,周东平,卢瑛,黄荣春..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):5276-5286,11.基金项目
广西农业科技自筹经费项目(Z2024128) （Z2024128）
国家自然科学基金(32160788) （32160788）
