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Related Experiment Videos

Semliki Forest virus as an expression vector in insect cell lines.

M M Pettigrew1, S L O'Neill

  • 1Section of Vector Biology, Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, CT 06510, USA.

Insect Molecular Biology
|September 1, 1999
PubMed
Summary

Replication-deficient Semliki Forest virus vectors efficiently express foreign genes in multiple insect cell lines, including Aedes albopictus and Aedes aegypti. Novel viral transcripts were detected in some cell lines, indicating successful viral transcription and potential for further study.

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Area of Science:

  • Molecular Biology
  • Virology
  • Entomology

Background:

  • Semliki Forest virus (SFV) is a widely used alphavirus.
  • Expression vectors are crucial tools for gene expression studies.
  • Insect cell lines offer a valuable system for studying viral replication and gene expression.

Purpose of the Study:

  • To evaluate the efficacy of replication-deficient Semliki Forest virus (SFV) expression vectors for foreign gene expression in various insect cell lines.
  • To quantify the levels of foreign gene product (GFP) in infected insect cells.
  • To analyze viral transcription patterns in different insect cell lines following SFV vector infection.

Main Methods:

  • Infection of multiple insect cell lines (Aedes albopictus, Aedes aegypti, Anopheles gambiae) with SFV-GFP expression vectors.

Related Experiment Videos

  • Quantification of green fluorescent protein (GFP) expression using fluorescence microscopy or other methods.
  • Northern blot analysis to detect and characterize viral transcripts.

    • Main Results:

    • Nearly 100% infection rates were achieved in Aedes albopictus (C6/36, Aa23T) and Aedes aegypti (MOS20) cell lines.
    • SFV vectors successfully infected an Anopheles gambiae cell line (MOS55).
    • Novel viral transcripts were identified in Aa23T, C6/36, and MOS55 cell lines, but not in BHK or MOS20 cells.

    Conclusions:

    • Replication-deficient SFV expression vectors are effective tools for high-level foreign gene expression in diverse mosquito cell lines.
    • SFV infection leads to distinct viral transcription profiles in different insect cell lines.
    • These findings support the use of SFV vectors for genetic studies in medically important insect vectors.