Expression And Identification Of The HCV Core Protein In The Human Hepatocyte QSG7701

Objective To construct a recombinant plasmid enconding HCV core gene and induce theexpression of core protein in the human hepatocyte and identify the core protein.Method HCV core gene was clone by using PCR from plasmid pBRTM/HCV1-3310 whichconcludes the full length of HCV gene. We recombined the core segment with expressionplasmid pcDNA3.1(-) to construct eukaryotic plasmid pcDNA3.1(-)/core. Then the plasmidpcDNA3.1(-)/core was transfected into human hepatocytes by using poly-cation. Theexpression of core protein was detected by immunochemical staining method and Westernblot.Result 1 The clone core segment had correct length and sequence .2 The QSG7701 cells were successfully transfected by the eukaryotic expressed plasmid pcDNA3.1(-)/core and screened by G418.3 Core protein were expressed in the QSG7701 and its relevant molecular weight was 21KD detected by. immunohistochemical staining and Western blotting.Conclusion We have successfully constructed eukaryotic expression plasmid pcDNA3.1(-)/core which included the HCV core gene. And the vector successfully expressed core protein in human hepatocytes . these result provide a good core protein expression system for the study of the hepatocarcinogesis of the core protein and lay foundation for developingthe HCV DNA vaccine.