Abstract: Objective To construct a recombinant eukaryotic expression plasmid N2ICD/pCMV-Tag4 and transfect it into HEK 293T cells.Methods Human Notch2 intracellular domain gene(N2ICD) from Hela cells was amplified by reverse transcription-PCR(RT-PCR) and identified by enzyme restriction, sequencing and blasting and then cloned into eukaryotic expression vector pCMV-Tag4 to construct N2ICD/pCMV-Tag4, which was then transiently and stably transfected into HEK 293T cells.Quantitative-PCR(Q-PCR) and Western blot was used to detect target protein expression.Results N2ICD was obtained by RT-PCR.The recombinant eukaryotic expression vector Notch2 intracellular domain(N2ICD)/pCMV-Tag4 was successfully constructed and N2ICD was expressed in HEK 293T cells.Conclusion N2ICD was expressed in HEK 293T cells, which provides a foundation for further investigating the function of Notch2 receptor.