Hepatitis C is an RNA virus that has infected around 170 million people worldwide. The characterization of this virus and the proteins that compose it has become a widely researched field. DExD/H-box helicase 58 (LGP2) is a dsRNA helicase that is involved in the Interferon I (IFN-1) pathway which begins the antiviral response of cells upon viral infection. This project aimed to create fluorescent versions of the cellular helicase LGP2 in a eukaryotic expression system. A yellow fluorescent protein (YFP) was successfully attached to LGP2. The plasmid construct pCMV-mYFP-LGP2 was successfully expressed in HEK293T cells and observed under a fluorescence microscope. Although the cells expressing the protein were killed, fluorescence could be attributed to the YFP portion of the protein based on the experimental design. It is theorized that the level of expression resulting from the experimental conditions was higher than physiological levels and this led to cell death.

Design and cloning of fluorescent human helicase DHX58 (LGP2): Building the tools to study viral helicases

247th National Meeting (2014)

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