畜牧与兽医2025，Vol.57Issue(5)：15-23,9.
鸡SOX6基因真核表达载体构建及其对成肌细胞分化相关基因表达的影响
Construction of eukaryotic expression vector of chicken SOX6 gene and its effect on gene expression of myoblast differentiation
摘要
Abstract
This study was to investigate the temporal and spatial expression of chicken SOX6 gene and its role in myoblast differentiation.Fluorescence quantitative PCR was used to detect the expression of SOX6 mRNA in different tissues of chickens as well as in skeletal muscles during embryonic stage(E11-E19)and early post-hatching period(D0,D3,D6).The coding sequence region(CDS)of the SOX6 gene was amplified and cloned by RT-PCR.After sequencing,the physicochemical properties and structural functions of the SOX6 protein were predicted by bioinformatics tools.The eukaryotic expression vector of the SOX6 gene was constructed and transfected into the primary myo-blasts in the chickens,and the recombinant protein was detected by Western blot.Finally,the effect of the overexpression and knockdown of the SOX6 gene on the expression of myogenic differentiation-related genes was detected by fluorescence quantitative PCR.The results showed that the SOX6 mRNA was expressed in 12 various tissues of the chickens,with higher expression in the pectoral muscle and leg muscle,and lower expression in the stomach and spleen.The expression of SOX6 mRNA gradually increased during the embryonic stage from E1 1 to E19,and was significantly higher in the D6 stage than in the other stages(P＜0.05).The CDS of the chicken SOX6 gene was obtained by RT-PCR.The total length of the coding region of the chicken SOX6 gene was 2 367 bp,encoding 789 amino acids.The bioinformatics analysis showed that the chicken SOX6 protein was a neutral,hydrophilic protein,with a molecular weight of 87.533 kDa,an isoelectric point of 6.99,instability coefficient of 60.95,and fat coefficient of 62.51.The SOX6 overexpression vector was constructed and transfected into chicken primary myoblasts,and SOX6-MYC fusion protein was successfully expressed.After SOX6 was overexpressed in the myoblasts,the mRNA expressions of the MyoD and MyoG genes increased significantly(P＜0.05),while the MyoD and MyoG mRNA expressions were sig-nificantly decreased after knocking down SOX6(P＜0.05).The results indicated that the SOX6 gene played a regulatory role in the differen-tiation and development of the myoblasts.The results of this study lay a foundation for further analysis of the function of the SOX6 gene.关键词
鸡/SOX6基因/克隆/真核表达/成肌细胞分化Key words
chicken/SOX6 gene/cloning/eukaryotic expression/myoblast differentiation分类
畜牧业引用本文复制引用
刘书鸣,魏志恒,彭鑫,周博,夏磊,裴军,徐璐,郁建峰,顾志良..鸡SOX6基因真核表达载体构建及其对成肌细胞分化相关基因表达的影响[J].畜牧与兽医,2025,57(5):15-23,9.基金项目
江苏省自然科学基金项目(BK20191476) （BK20191476）
江苏省大学生创业创新重点项目(202310333012Z) （202310333012Z）
