山西医科大学学报2024，Vol.55Issue(7)：849-854,6.DOI:10.13753/j.issn.1007-6611.2024.07.006
小鼠SGK3真核表达载体的构建及鉴定
Construction and identification of mouse SGK3 eukaryotic expression vector
摘要
Abstract
Objective To construct the eukaryotic expression vector pcDNA3.1-MYC-SGK3-mCherry containing mouse serum and glucocorticoid-induced protein kinase 3(SGK3)gene,and verify its expression in HEK293 cells after transfection.Methods The target gene SGK3 in the eukaryotic expression plasmid pcDNA3.1-MYC-SGK3 preserved in the laboratory was fused with mCherry and amplified by polymerase chain reaction,and then cloned into pcDNA3.1-MYC plasmid.After restriction enzyme digestion and sequenc-ing,it was transfected into HEK293 cells by liposome method,and the protein expression of the target gene was detected by Western blotting.Results The sequencing results were consistent with the previous expected results,confirming that the pcDNA3.1-MYC-SGK3-mCherry eukaryotic expression vector was successfully constructed.Western blotting results showed that there was clear positive reaction band in HEK293 cells transfected with pcDNA3.1-MYC-SGK3-mCherry,indicating that the target fragment was successfully expressed.Conclusion The eukaryotic expression vector pcDNA3.1-MYC-SGK3-mCherry is successfully constructed.关键词
血清和糖皮质激素诱导蛋白激酶3/真核表达载体/聚合酶链式反应/限制性内切酶/HEK293细胞/载体构建Key words
serum and glucocorticoid induced protein kinase 3/eukaryotic expression vector/polymerase chain reaction/restriction endonuclease/HEK293 cells/vector construction分类
生物科学引用本文复制引用
巴隆,张丽娜,孟峻..小鼠SGK3真核表达载体的构建及鉴定[J].山西医科大学学报,2024,55(7):849-854,6.基金项目
国家自然科学基金资助项目(81360109,81660267) （81360109,81660267）
内蒙古自治区科技厅自然科学基金资助项目(2021MS08158) （2021MS08158）
