Related Experiment Videos
Development and application of a reverse genetics system for Japanese encephalitis virus
Sang-Im Yun1, Seok-Yong Kim, Charles M Rice
1Department of Internal Medicine, Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Korea.
Journal of Virology
|May 14, 2003
Summary
Researchers developed a stable Japanese encephalitis virus (JEV) cDNA clone for genetic studies. This infectious clone facilitates JEV research and potential vaccine development.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Japanese encephalitis virus (JEV) causes significant mortality and morbidity, particularly in children.
- Molecular genetic studies of JEV are limited by the absence of a stable, full-length infectious cDNA clone.
Purpose of the Study:
- To develop a genetically stable, full-length infectious JEV cDNA clone.
- To establish a novel reverse genetics system for JEV research.
Main Methods:
- Sequencing of a JEV isolate and cloning its full-length cDNA into a bacterial artificial chromosome.
- Engineering the clone for in vitro transcription of synthetic RNAs with authentic ends.
- Infecting susceptible cells with synthetic RNAs and characterizing recovered viruses.
Main Results:
- Produced highly infectious synthetic RNAs yielding high titers of synthetic JEV.
- Recovered JEV was indistinguishable from the parental virus in multiple biological characteristics.
- Demonstrated cDNA stability in E. coli and successful genetic manipulation via a marker mutation.
Conclusions:
- The developed JEV cDNA clone and reverse genetics system significantly advance JEV research.
- This system is valuable for studying JEV biology and developing JEV vaccines.
- JEV shows potential as a heterologous gene expression vector.