A protocol to express and isolate HRV16 3C protease for use in protease assays

Erin WALKER, Reena GHILDYAL

Research output: A Conference proceeding or a Chapter in BookChapter

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.

Original languageEnglish
Title of host publicationRhinoviruses
Subtitle of host publicationMethods and Protocols
EditorsD.A Jans, R Ghildyal
Place of PublicationUSA
PublisherHumana Press
Pages143-148
Number of pages6
Volume1221
ISBN (Print)9781493915705
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745

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Rhinovirus
Peptide Hydrolases
Bacterial Transformation
Plasmids
Complementary DNA
Serogroup
3C proteases
In Vitro Techniques

Cite this

WALKER, E., & GHILDYAL, R. (2015). A protocol to express and isolate HRV16 3C protease for use in protease assays. In D. A. Jans, & R. Ghildyal (Eds.), Rhinoviruses: Methods and Protocols (Vol. 1221, pp. 143-148). (Methods in Molecular Biology). USA: Humana Press. https://doi.org/10.1007/978-1-4939-1571-2_11
WALKER, Erin ; GHILDYAL, Reena. / A protocol to express and isolate HRV16 3C protease for use in protease assays. Rhinoviruses: Methods and Protocols. editor / D.A Jans ; R Ghildyal. Vol. 1221 USA : Humana Press, 2015. pp. 143-148 (Methods in Molecular Biology).
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WALKER, E & GHILDYAL, R 2015, A protocol to express and isolate HRV16 3C protease for use in protease assays. in DA Jans & R Ghildyal (eds), Rhinoviruses: Methods and Protocols. vol. 1221, Methods in Molecular Biology, Humana Press, USA, pp. 143-148. https://doi.org/10.1007/978-1-4939-1571-2_11

A protocol to express and isolate HRV16 3C protease for use in protease assays. / WALKER, Erin; GHILDYAL, Reena.

Rhinoviruses: Methods and Protocols. ed. / D.A Jans; R Ghildyal. Vol. 1221 USA : Humana Press, 2015. p. 143-148 (Methods in Molecular Biology).

Research output: A Conference proceeding or a Chapter in BookChapter

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KW - Recombinant protein purification

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WALKER E, GHILDYAL R. A protocol to express and isolate HRV16 3C protease for use in protease assays. In Jans DA, Ghildyal R, editors, Rhinoviruses: Methods and Protocols. Vol. 1221. USA: Humana Press. 2015. p. 143-148. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-1571-2_11