Construction and expression of the recombinant plasmid pEGFP-N1-Mxi1-0

Objective:To construct the recombinant eukaryote expression vector containing Max interacting protein(Mxi)1-0 gene,and detect the expression of Mxi1-0 in mouse embryo fibroblast(NIH/3T3) cells for providing the basis to explore the effect and mechanism of Mxi1-0.Methods:Mxi1-0 gene was cloned by RT-PCR from cancer cells,which was subcloned into enhanced green fluorescent protein eukaryote expression vector(pEGFP-N1) to construct the recombinant vector pEGFP-N1-Mxi1-0.The recombinant vector pEGFP-N1-Mxi1-0 was identified by enzyme digestion and sequencing,which was transfected into the NIH/3T3 cells by lipidosome.The protein expression of Mxi1-0 in NIH/3T3 cells was detected by Western blot,the intracellular localization of Mxi1-0 was investigated by immunofluorescence.Results:The recombinant eukaryote expression vector encoding Mxi1-0 was successfully constructed.The expression of Mxi1-0 in NIH/3T3 cells could be detected by Western blot.The Mxi1-0 localized in the cytoplasm of NIH/3T3 cells.Conclusions:The recombinant expression vector pEGFP-N1-Mxi1-0 is successfully constructed.The Mxi1-0 expression in NIH/3T3 cells can be detected,which localizes in the cytoplasm.