Construction and expression of eukaryotic expression vector containing Hantaan virus S gene

  Objective   Hantaan virus S gene encoding Nucleocapsid protein was cloned into the eukaryotic expressing plasmid pTARGET^TM to get the recombinant plasmid pTARGET-hans, then to express it in vitro.

  Methods   Polymerase chain reaction(PCR) was used to clone S gene segment and its product was inserted into eukaryotic expression vector pTARGET^TM; then the recombinant expression vector pTARGET-hans was identified by restriction enzyme analysis and transfected into Vero-E6 cells by electroporation.The transient expression of Hantaan virus nucleocapsid protein in Vero-E6 cells was detected by indirect immunofluorescence assay (IFA).

  Results   Restriction enzyme analysis demonstrated that S gene was successfully inserted into pTARGET^TM and IFA result showed that green fluorescence was observed in some transfected Vero-E6 cells.

  Conclusion   The eukaryotic expression vector pTARGET-hans was successfully constructed and expressed in vitro, which would lay a foundation for further researching S gene segment immunization against Hantaan Virus.