CONSTRUCTION OF pcDNA3.1-preS1-GFP RECOMBINANT PLASMID AND EXPRESSION OF SPERM IN MICE

Objective To construct pcDNA3.1-preS1-GFP recombinant eukaryotic expression vector harboring green fluorescent protein(GFP)and hepatitis B virus(HBV)preS1 gene and observe the expression of sperm in mice.Methods The HBV preS1 gene fragment was amplified from the HBV genome plasmid by PCR and inserted into eukaryotic expression vector pcDNA3.1-GFP.After the identification by digestion and sequencing,the recombinant eukaryotic expression vector was transfec-ted into sperm of mouse by Lipofectamine TM2000.Expression of GFP was examined directly with fluorescence microscope.The existence and expression of HBV preS1 gene was detected by PCR.Results The eukaryotic expression vector was created successfully and transfected to sperm of mouse with pcDNA3.1-preS1-GFP recombinant plasmid,green fluorescence was observed with fluorescence microscope,which presented on the head of sperm,and a specific band at 589 bp tha in the area of HBV preS1 gene was obtained by PCR.Conclusion The successful construction of the pcDNA3.1-preS1-GFP recombinant plasmid and its expression in sperm of mouse establishes an experiment foundation for creating a new mouse model of vertical-transmission HBV DNA.