小鼠SGK3真核表达载体的构建及鉴定OACSTPCD

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.