Presenter(s)

    Sara M. Hubbell

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    Description

    Preliminary data estimates there were about 60,000 deaths and 800,000 hospitalizations due to the influenza virus for the 2017-2018 season (CDC.gov). With the ultimate goal of alieving some of this disease burden, current research in our lab focuses on developing a new antiviral treatment for influenza, since there are very limited options for battling influenza infection besides seasonal vaccination and non-ideal medications like Tamiflu. The Polymerase Acidic (PA) influenza protein is an appealing target for drug design due to its low genetic drift and requirement for influenza replication. The first step in designing a drug to inhibit PA is to isolate and purify a soluble version of the protein to enable structure-based drug design. Recombinant PA expressed in bacteria in our lab has demonstrated very low solubility. We hypothesized that by protecting a hydrophobic binding site, a fusion of PA and its natural binding partner, Polymerase Basic protein 1 (PB1), would be more soluble than PA alone. Mutagenic PCR primers were designed and used to attach the sequence coding for a PB1 peptide to the 5’ and 3’ ends of PA’s coding sequence, resulting in two different protein fusions in cloning vectors. Golden Gate Assembly was then used to insert this recombinant DNA into expression vectors containing fusion partners to increase the solubility of PA even further. A collection of eight expression vectors with varying protein fusions have been successfully designed and cloned. Small-scale solubility testing of these constructs is underway.

    Publication Date

    4-22-2021

    Project Designation

    Independent Research

    Primary Advisor

    Douglas S. Daniels

    Primary Advisor's Department

    Chemistry

    Keywords

    Stander Symposium project, College of Arts and Sciences

    United Nations Sustainable Development Goals

    Good Health and Well-Being

    Design and Creation of PB1-Peptide Fusions to Solubilize Recombinant Influenza PA Protein

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