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Related Experiment Videos

Recombinant replication-restricted VSV as an expression vector for murine cytokines.

Mark A Miller1, Christy L Lavine, Sheri D Klas

  • 1Department of Molecular Sciences, University of Tennessee Health Science Center, 858 Madison Avenue, Memphis, TN 38163, USA. mamiller@utmem.edu

Protein Expression and Purification
|December 19, 2003
PubMed
Summary

Recombinant vesicular stomatitis virus (VSV) systems efficiently produce large amounts of bioactive murine cytokines. This deltaG-VSV platform offers a rapid and cost-effective method for cytokine expression and bioactivity assessment.

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Area of Science:

  • Virology
  • Molecular Biology
  • Immunology

Background:

  • Vesicular stomatitis virus (VSV) is a negative-strand RNA virus known for its ability to rapidly inhibit host protein synthesis.
  • VSV's protein production machinery can be engineered to express high levels of viral proteins, including foreign gene products.
  • Recombinant VSV systems offer a powerful tool for protein expression and manipulation.

Purpose of the Study:

  • To develop a recombinant VSV system for efficient expression of murine cytokines.
  • To utilize a deltaG-VSV strategy for enhanced protein secretion and bioactivity.
  • To assess the quantity and bioactivity of cytokines produced using this novel expression system.

Main Methods:

  • Creation of recombinant deltaG-VSV encoding various murine cytokines using reverse genetics.

Related Experiment Videos

  • Propagation of deltaG-VSV in G protein-expressing cells to generate complemented virions.
  • Infection of BHK cells with deltaG-VSV constructs to monitor and measure cytokine production.
  • Assessment of the bioactivity of the expressed cytokines.

    • Main Results:

    • deltaG-VSV constructs successfully expressed high levels of recombinant murine cytokines.
    • The deltaG-VSV system demonstrated efficient protein expression and secretion.
    • The produced cytokines were found to be bioactive, indicating functional protein production.
    • This method provides a rapid and cost-effective means for cytokine generation.

    Conclusions:

    • The deltaG-VSV system is a highly effective platform for the rapid and inexpensive production of large quantities of bioactive murine cytokines.
    • This approach leverages VSV's inherent protein expression capabilities for biotechnological applications.
    • The developed system has significant potential for research and therapeutic applications involving cytokines.