Construction of RNAi eukaryotic expression vector pSilencer31-H1/GFP a nd its expression in HCT-8 cells

Objective To construct the eukaryotic expression plasmid containing green fluorescent prot ein gene and observe its expression in HCT-8 cells,which lays the foundation fo r further development of RNAi related research.Method GFP was obtained from pEGFP-C1 by PCR technology and was inserted into pSilencer3.1-H1.After identified by restriction endonucleas e and sequence analysis,the recombinant expression plasmid was transfected with l ipofectamine 2000 to HCT-8 cells.The expression was observed under a fluorescen t microscope.Results Restr i ctive endonuclease assay and sequence analysis verified the successful construct ion of the recombinant vector pSilencer3.1-H1/GFP,and GFP protein was stable e x pressed in HCT-8 cells.Conclusion The GFP eukaryotic expression plasmid is successfully constructed and ex pressed in HCT-8 cells.