Construction of Eukaryotic Fluorescence Expression Vector of Rat p75NTR and Its Expression in HEK293 cells

Objective:To construct a eukaryotic fluorescence expression vector of human p75 neurotrophin receptor(p75NTR) cDNA and investigate its expression in human embryo kidney 293(HEK293) cells.Methods:The pEGFP-N1-RP75 eukaryotic fluores-cence expression vector was constructed by cloning a 828bp insertion of rat p75NTR cDNA from pDC316-RP75 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 was transfected into HEK293 cells by lipofection and the expression of rat p75NTR in HEK293 was detected by confo-cal microscopy and immunohistochemistry staining.Results:The analysis of enzyme hydrolyze and DNA sequence showed that the re-combinant eukaryotic expression vector contained rat p75NTR cDNA and the detection of confocal microscopy and immunohistochem-istry staining showed that the HEK293 cells transfected with pEGFP-N1-RP75 could express rat p75NTR.Conclusions:The results showed that the recombinant eukaryotic fluorescence expression vector was constructed successfully and the transfected HEK293 cells could express rat p75NTR,which paves the way for further studies on the mechanism of p75NTR function.