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Cloning and expression of bovine herpesvirus-1 glycoprotein C
P K Gupta1, M Saini, L K Gupta
1National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar, India.
Insights
Researchers expressed Bovine Herpesvirus-1 (BHV-1) Glycoprotein C (gC) in mammalian cells. This surface expression of gC in MDBK cells is a key step for further vaccine development against BHV-1.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Glycoprotein C (gC) of Bovine Herpesvirus-1 (BHV-1) is crucial for viral infection and immunity.
- gC is highly expressed on infected cells and the virus envelope, eliciting a protective antibody response in cattle.
Purpose of the Study:
- To express BHV-1 gC in mammalian cells for further study.
- To investigate the feasibility of using recombinant DNA technology for gC expression.
Main Methods:
- The gC gene fragment was cloned into a plasmid under the control of the RSV-LTR promoter.
- The resultant plasmid (pRSV-gC) was transfected into MDBK cells.
- Indirect immunofluorescence was used to detect gC expression on the cell surface.
Main Results:
- Successful expression of BHV-1 gC was achieved in MDBK cells.
- Non-permeabilized cells showed surface expression of gC, indicating proper folding and localization.
- The RSV-LTR promoter effectively drove gC expression.
Conclusions:
- Mammalian cells can be successfully engineered to express functional BHV-1 gC.
- This expression system provides a platform for studying gC's role in viral pathogenesis and for developing subunit vaccines.
- Surface expression of gC is achievable, which is vital for eliciting a protective immune response.
Abstract:
Glycoprotein C (gC) of Bovine Herpesvirus-1 (BHV-1) is expressed at high levels on surface of infected cells and on virus envelope. It is relatively immunodominant in antibody response to BHV-1 infection and protective in immunized bovines against BHV-1 challenge. In an attempt to express gC in mammalian cells, the 2.4 kb BamHI-EcoRI fragment, containing complete coding sequence of the gC gene was excised from a recombinant plasmid and cloned under the control of RSV-LTR. The resultant plasmid pRSV-gC was transfected into MDBK cells and expression of gC was detected by indirect immunofluorescence. The non-permeabilized cells revealed surface expression of gC.
