[Construction of mammalian cell lines continuously expressing JEV structural proteins]

Area of Science:

• Virology
• Molecular Biology
• Vaccine Development

Context:

• Japanese encephalitis virus (JEV) poses a significant public health challenge.
• Developing stable cell lines for JEV research is crucial for vaccine and therapeutic advancements.
• Previous attempts using JEV prM-E genes resulted in cell toxicity.

Purpose:

• To establish stable cell lines expressing JEV structural proteins for easier production of virus-like particles.
• To investigate the role of the JEV capsid protein in cell line stability.
• To compare the expression of prM-E genes versus C-prM-E genes in mammalian cells.

Summary:

• Recombinant plasmids containing JEV prM-E and C-prM-E genes were constructed.
• Transfection of BHK-21 cells with pCJE-ME (prM-E) led to cell death.
• Transfection with pCJE-CME (C-prM-E) resulted in stable cell lines selected by G418, confirmed by ELISA, Western Blot, and IFA.
• The JEV capsid protein (from C-prM-E) enhanced the stability of cell lines expressing JEV structural proteins.

Impact:

• Successfully established stable JEV-expressing cell lines using the C-prM-E construct.
• Demonstrated the stabilizing effect of the JEV capsid protein on these cell lines.
• Facilitated easier acquisition of JEV virus-like particles for further study and potential applications.