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Rhinoviruses pp 143-148 | Cite as

A Protocol to Express and Isolate HRV16 3C Protease for Use in Protease Assays

  • Lora M. Jensen
  • Erin J. Walker
  • Reena GhildyalEmail author
Protocol
  • 2.1k Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 1221)

Abstract

Human rhinovirus (HRV) proteases are highly conserved across serotypes and have very similar target specificity. However, there are some serotype-specific differences in their action. It is therefore necessary when performing in vitro protease assays to ensure that the recombinant proteases are specific to the serotype of the HRV under study. We describe a simple method for isolating HRV16 3C protease from a bacterial expression system, including transformation of bacterial cells with a commercially available cDNA plasmid which can be adapted to use for 3C proteases from any other HRV serotypes. The extracted, active 3C protease can then be used for in vitro protease assays.

Key words

HRV16 3C protease In vitro protease activity assays Recombinant protein purification 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Lora M. Jensen
    • 1
  • Erin J. Walker
    • 1
  • Reena Ghildyal
    • 1
    Email author
  1. 1.Faculty of Education, Science, Technology and Mathematics, Centre for Research in Therapeutic SolutionsUniversity of CanberraCanberraAustralia

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