Construction of Eukaryotic Fluorescence Expression Plasmid of Human P75NTR and Its Expression in HEK293 Cells

1.Objective: To establish a eukaryotic fluorescence expression plasmid of human p75 neurotrophin receptor(p75NTR) cDNA and investigate its expression in human embryo kidney 293(HEK293) cells.2.Methods: The pEGFP-N1-HP75 eukaryotic fluorescence expression vector is constructed by cloning a 835bp insertion of human p75NTR cDNA from pDC316-HP75 plasmid containing the full length open reading frame to pEGFP-N1 vector.After analysis of enzyme hydrolyze and DNA sequence,the recombinant eukaryotic expression vector is transfected into HEK293 cells by lipofection and the expression of human p75NTR in HEK293 is detected by confocal microscopy and Western blot.3.Results: The analysis of enzyme hydrolyze and DNA sequence shows that the recombinant eukaryotic expression vector contained human p75NTR cDNA and the detection of confocal microscopy and Western blot show that the HEK293 cells transfected with pEGFP-N1-HP75 could express rat p75NTR.4.Conclusion: The recombinant eukaryotic fluorescence expression vector is constructed successfully and the transfected HEK293 cells could express human p75NTR,which paves the way for further studies on the mechanism of p75NTR function.