饲料研究2025，Vol.48Issue(9)：64-69,6.DOI:10.13557/j.cnki.issn1002-2813.2025.09.012
蛋鸡CCND1基因真核表达载体的构建及在卵泡不同发育时期的时空表达模式研究
Construction of eukaryotic expression vector of CCND1 gene in laying hens and its spatiotemporal expression pattern in different developmental stages of follicles
摘要
Abstract
The experiment was to explore the spatiotemporal expression pattern of cyclin D1(CCND1)gene in laying hens follicles at different development stages and delve into its molecular biological functions.A eukaryotic expression vector has been constructed to predict CCND1's specific molecular roles during laying hens follicle development.Follicles of varying sizes from 150-day-old Hyline brown laying hens were selected as experimental materials.Semi-quantitative RT-PCR and immunohistochemistry were used for molecular-level spatiotemporal expression analysis.After verifying the ligation product's correctness via double enzyme digestion with BamH Ⅰ and XhoⅠ,sequencing was performed.The correctly sequenced eukaryotic expression vector was then transfected into chicken ovarian granulosa cells,and the transfection efficiency is confirmed using real-time fluorescent quantitative PCR.The results showed that CCND1 mRNA expression was significantly higher in follicles with diameters of 5～6 mm and 6～7 mm than in other follicles(P＜0.05).CCND1 protein localization was observed in oocytes and theca of pre-hierarchical follicles.A eukaryotic expression vector pYr-adshuttle-4-CCND1 had been successfully constructed through colony PCR amplification and sequencing analysis and effectively transfected into chicken follicle granulosa cells.The study indicates that CCND1 likely plays a role in laying hens follicle growth and development,offering a vital molecular basis and tool for future gene-function and application studies.关键词
蛋鸡/细胞周期蛋白D1(CCND1)/卵泡/真核表达载体/时空表达Key words
laying hens/CCND1/follicle/eukaryotic expression vectors/spatiotemporal expression分类
农业科技引用本文复制引用
邹贤群,余静,刘永杰,陈晓霞..蛋鸡CCND1基因真核表达载体的构建及在卵泡不同发育时期的时空表达模式研究[J].饲料研究,2025,48(9):64-69,6.基金项目
吉林农业科技学院国家级大学生创新项目(项目编号:202311439013) （项目编号:202311439013）
