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

Erin WALKER; Reena GHILDYAL

doi:10.1007/978-1-4939-1571-2_11

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

Research output: A Conference proceeding or a Chapter in Book › Chapter

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 language	English
Title of host publication	Rhinoviruses
Subtitle of host publication	Methods and Protocols
Editors	D.A Jans, R Ghildyal
Place of Publication	USA
Publisher	Humana Press
Pages	143-148
Number of pages	6
Volume	1221
ISBN (Print)	9781493915705
DOIs	https://doi.org/10.1007/978-1-4939-1571-2_11
Publication status	Published - 2015

Publication series

Name	Methods in Molecular Biology
ISSN (Print)	1064-3745

Access to Document

10.1007/978-1-4939-1571-2_11

Cite this

@inbook{ef0eeebf3d4d4deb8f9a8a4e45f24ed3,

title = "A protocol to express and isolate HRV16 3C protease for use in protease assays",

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.",

keywords = "HRV16 3C protease, In vitro protease activity assays, Recombinant protein purification",

author = "Erin WALKER and Reena GHILDYAL",

year = "2015",

doi = "10.1007/978-1-4939-1571-2_11",

language = "English",

isbn = "9781493915705",

volume = "1221",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "143--148",

editor = "D.A Jans and R Ghildyal",

booktitle = "Rhinoviruses",

address = "United States",

}

TY - CHAP

T1 - A protocol to express and isolate HRV16 3C protease for use in protease assays

AU - WALKER, Erin

AU - GHILDYAL, Reena

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - HRV16 3C protease

KW - In vitro protease activity assays

KW - Recombinant protein purification

UR - http://www.scopus.com/inward/record.url?scp=84921720404&partnerID=8YFLogxK

UR - http://www.mendeley.com/research/protocol-express-isolate-hrv16-3c-protease-protease-assays

U2 - 10.1007/978-1-4939-1571-2_11

DO - 10.1007/978-1-4939-1571-2_11

M3 - Chapter

SN - 9781493915705

VL - 1221

T3 - Methods in Molecular Biology

SP - 143

EP - 148

BT - Rhinoviruses

A2 - Jans, D.A

A2 - Ghildyal, R

PB - Humana Press

CY - USA

ER -

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

Abstract

Publication series

Access to Document

Other files and links

Fingerprint

Cite this