Creating a Stable Marker Virus Using EGFP in Semliki Forest Virus
Author Information
Author(s): Nele Tamberg, Valeria Lulla, Rennos Fragkoudis, Aleksei Lulla, John K. Fazakerley, Andres Merits
Primary Institution: Estonian Biocentre, Tartu, Estonia
Hypothesis
Can the insertion of an enhanced green fluorescent protein (EGFP) into the replicase gene of Semliki Forest virus create a genetically stable marker virus?
Conclusion
The study successfully created a genetically stable marker virus that expresses EGFP and maintains neurovirulence.
Supporting Evidence
- The engineered virus was able to replicate to high titres and remained neurovirulent.
- Over 90% of the viruses remained EGFP-positive after five passages in vitro.
- Only one EGFP-negative plaque was observed after five in vivo passages.
Takeaway
Scientists made a virus that glows green under a special light, which helps them study how the virus works without it changing too much.
Methodology
The virus was engineered using subcloning and PCR-based mutagenesis, followed by infectious-centre assays and one-step growth studies.
Limitations
The genetic stability of the virus may still be compromised over multiple passages, as some genomes with deletions were observed.
Participant Demographics
5–6-week-old female BALB/c mice were used for in vivo studies.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website