Abstract
The endonuclease activity of influenza A virus RNA polymerase allows the digestion of host mRNA. The PA endonuclease domain could be a target of anti-influenza A virus drugs such as glycoconjugates. To test this activity, purified viral PA endonuclease domain protein is necessary. Here, we describe a method for the expression and purification of recombinant influenza A virus PA endonuclease domain protein, and a PA endonuclease assay to test glycoconjugates for potential inhibitory activity. Using influenza A virus PA cDNA as a template, DNA from the open reading frame of the PA endonuclease domain protein was amplified with PCR and cloned into an expression vector. Six His-tagged PA endonuclease domain proteins were expressed in Escherichia coli and purified with Ni2+-agarose resin and imidazole using an ion-exchange column. Using the recombinant PA endonuclease domain protein, an endonuclease assay was performed.
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References
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Acknowledgment
This work was supported by Grants-in-Aid for Scientific Research (C) Number 22590422 and 25460574 to T.K.
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Kuzuhara, T. (2022). Preparation of Recombinant PA Endonuclease Domain Protein of Influenza A Virus and Its Application for Glycobiology Research. In: Suzuki, Y. (eds) Glycovirology. Methods in Molecular Biology, vol 2556. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2635-1_7
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DOI: https://doi.org/10.1007/978-1-0716-2635-1_7
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