Objective:This study aimed to prepare Golgi protein-73(GP73)-DC vaccine by constructing human lentiviral vector of GP73and transfecting this vector in vitro into a murine dendritic cell line DC 2.4.This will lay the f...Objective:This study aimed to prepare Golgi protein-73(GP73)-DC vaccine by constructing human lentiviral vector of GP73and transfecting this vector in vitro into a murine dendritic cell line DC 2.4.This will lay the foundation for further study on targeted therapy of hepatocellular carcinoma using GP73-DC vaccine.Methods:Human GP73 gene was cloned into lentivirus GV358vector and validated by polymerase chain reaction(PCR)and sequencing.The validated GV358-GP73vector was then cotransfected with pHelper 1.0plasmid into 293Tcells,in order to prepare lentivirus GP73vector.Finally,the protein expression of GP73 was achieved by transfecting the lentivirus GP73vector into DC 2.4cells in vitro and determined by Western blotting.Results:PCR and sequencing confirmed that the GV358-GP73lentivirus vector was constructed correctly,and the titer level reached 2×108TU/mL.After transfection in DC 2.4cells,the GP73protein level was significantly enhanced.Conclusion:The lentivirus GV538-GP73vector was constructed successfully,and could be effectively expressed in DC 2.4cells.展开更多
基金supported by the National Natural Science Foundation of China (No.81360347)the Key University and College Project of Guangxi Provisional Department of Education(No.ZD2014027).
文摘Objective:This study aimed to prepare Golgi protein-73(GP73)-DC vaccine by constructing human lentiviral vector of GP73and transfecting this vector in vitro into a murine dendritic cell line DC 2.4.This will lay the foundation for further study on targeted therapy of hepatocellular carcinoma using GP73-DC vaccine.Methods:Human GP73 gene was cloned into lentivirus GV358vector and validated by polymerase chain reaction(PCR)and sequencing.The validated GV358-GP73vector was then cotransfected with pHelper 1.0plasmid into 293Tcells,in order to prepare lentivirus GP73vector.Finally,the protein expression of GP73 was achieved by transfecting the lentivirus GP73vector into DC 2.4cells in vitro and determined by Western blotting.Results:PCR and sequencing confirmed that the GV358-GP73lentivirus vector was constructed correctly,and the titer level reached 2×108TU/mL.After transfection in DC 2.4cells,the GP73protein level was significantly enhanced.Conclusion:The lentivirus GV538-GP73vector was constructed successfully,and could be effectively expressed in DC 2.4cells.
