吉林大学学报（医学版）2025，Vol.51Issue(1)：51-57,7.DOI:10.13481/j.1671-587X.20250107
小鼠SGK1基因真核表达载体的构建及鉴定
Construction and identification of eukaryotic expression vector of mouse SGK1 gene
摘要
Abstract
Objective:To construct an eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry which containing mouse serum and glucocorticoid-induced kinase(SGK)1 gene,and to observe its expression in the transfected HEK293 cells.Methods:The SGK1 target gene segments were amplified by PCR method,and the segments were ligated to the pcDNA3.1-MYC-C-mcherry vector which was doubly-digested with Hind Ⅲ and Sbf Ⅰ.After successful verification by enzyme digestion and sequencing,the pcDNA3.1-MYC-SGK1-mcherry expression vector was transfected into the HEK293 cells by liposome transfection.Western blotting method was used to determine the expression level of eukaryotic expression vector in the cells.Results:The vector band was located at 5 200 bp and the target gene band was located at 3 100 bp,which was consistent with the expected results.The sequencing results were also consistent when compared with the expected sequence by Snap Gene software,which indicated that the eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry was successfully constructed.Successful expression of the eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry was observed by Western blotting method,in which the transfected cells showed well-defined bands near the relative molecular mass of 49 000.Conclusion:The eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry is successfully constructed,laying a solid foundation for the subsequent study on the transition mechanism of SGK1 gene in the early development of mouse fertilized egg cells.关键词
血清和糖皮质激素诱导激酶1/真核表达载体/HEK293细胞/载体构建/质粒Key words
Serum and glucocorticoid-induced kinase 1/Eukaryotic expression vector/HEK293 cells/Vector construction/Plasmids分类
生物学引用本文复制引用
张丽娜,巴隆,孟峻..小鼠SGK1基因真核表达载体的构建及鉴定[J].吉林大学学报（医学版）,2025,51(1):51-57,7.基金项目
国家自然科学基金项目(81360109,81660267) （81360109,81660267）
内蒙古自治区科技厅自然科学基金项目(2021MS08158) （2021MS08158）
