Related Experiment Video
Updated: Feb 25, 2026
Using Reverse Genetics to Manipulate the NSs Gene of the Rift Valley Fever Virus MP-12 Strain to Improve Vaccine Safety and Efficacy
Published on: November 1, 2011
Expression of domain III of the envelope protein from GP-78: a Japanese encephalitis virus
Sahil Kulkarni1, Sandeepan Mukherjee2, Krishna Mohan Padmanabha Das3
1Department of Zoonosis, Haffkine Institute for Training, Research and Testing, Acharya Donde Marg, Parel, Mumbai, 400 012 India.
Abstract:
Acute encephalitis caused by the Japanese encephalitis virus (JEV) represents a growing epidemic and is a cause for concern in Southeast Asia. JEV is transmitted to humans through the bite of the Culicine mosquito species. The virus genome comprising of an RNA strand also encodes the envelope protein (E) which surrounds the virus. The E protein aids in fusion of virus with the cellular membrane of the host cell with the help of three structurally distinct domains (DI, DII, DIII) that are connected by flexible hinge regions. Of these domains, DIII (JEV-DIII) has been reported to interact with the cellular membrane, aid viral entry and viral replication. Hence JEV-DIII has the potential to be an antigen that can provide immune protection to a JEV infection. In this study, we describe the cloning and expression of DIII of GP-78, a virulent strain of JEV prevalent in India. Our data clearly shows that JEV-DIII expressed from pVAC1 in HEK293T cells is membrane targeted. To our knowledge, this is the first demonstration of a recombinant construct that may block JEV entry into the cells and/or evoke specific antibodies against JEV. Future studies will reveal if our construct will elicit significant immune responses which will alleviate or ameliorate the pro-inflammatory responses induced by JEV.
Insights
Researchers engineered a Japanese encephalitis virus (JEV) envelope protein domain (JEV-DIII) to target cell membranes. This novel construct shows potential for blocking JEV entry and eliciting protective immune responses against the virus.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Japanese encephalitis virus (JEV) causes a significant epidemic of acute encephalitis in Southeast Asia.
- JEV is transmitted via *Culicine* mosquitoes, posing a public health concern.
- The JEV envelope (E) protein mediates viral entry through membrane fusion, with domain III (JEV-DIII) playing a key role.
Purpose of the Study:
- To clone and express the JEV-DIII from a virulent Indian strain (GP-78).
- To investigate the potential of JEV-DIII as a target for immune protection against JEV infection.
- To develop a recombinant construct for potential therapeutic or prophylactic applications.
Main Methods:
- Cloning of the JEV-DIII gene from the GP-78 strain.
- Expression of JEV-DIII in HEK293T cells using the pVAC1 vector.
- Analysis of the cellular localization and membrane targeting of the expressed JEV-DIII.
Main Results:
- Successful cloning and expression of JEV-DIII.
- Demonstrated membrane targeting of the expressed JEV-DIII.
- This is the first report of a recombinant construct potentially inhibiting JEV entry and/or inducing specific antibodies.
Conclusions:
- The engineered JEV-DIII construct is membrane-targeted, suggesting a potential mechanism to block viral entry.
- This recombinant JEV-DIII holds promise as a candidate antigen for eliciting protective immunity.
- Further research is needed to confirm its efficacy in inducing significant immune responses and mitigating JEV-induced inflammation.
