Reverse Genetic Engineering of the Human Rhinovirus Serotype 16 Genome to Introduce an Antibody-Detectable Tag

Erin WALKER, Joan Jensen, Reena GHILDYAL

Research output: A Conference proceeding or a Chapter in BookChapter

1 Citation (Scopus)

Abstract

The ability to accurately detect viral proteins during infection is essential for virology research, and the lack of specific antibodies can make this detection difficult. Reverse genetic engineering of virus genomes to alter the wild-type genome is a powerful technique to introduce a detectable tag onto a viral protein. Here we outline a method to incorporate an influenza hemagglutinin epitope tag onto the 2A protease of HRV16. The method uses site-directed mutagenesis PCR to introduce the sequence for the HA antigen onto either the C or N termini of 2A protease while keeping the relevant internal cleavage sites intact. The new viral product is then cloned into a wild-type HRV16 plasmid and transfected into Ohio Hela cells to produce recombinant virus.

Original languageEnglish
Title of host publicationRhinoviruses
Subtitle of host publicationMethods and Protocols
EditorsD.A Jans, R Ghildyal
Place of PublicationUSA
PublisherHumana Press
Pages171-180
Number of pages10
Volume1221
ISBN (Print)9781493915705
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745

Fingerprint

Reverse Genetics
Rhinovirus
Genetic Engineering
Viral Proteins
Peptide Hydrolases
Genome
Viruses
Virology
Antibodies
Hemagglutinins
Site-Directed Mutagenesis
HeLa Cells
Human Influenza
Epitopes
Plasmids
Antigens
Polymerase Chain Reaction
Infection
Research
Serogroup

Cite this

WALKER, E., Jensen, J., & GHILDYAL, R. (2015). Reverse Genetic Engineering of the Human Rhinovirus Serotype 16 Genome to Introduce an Antibody-Detectable Tag. In D. A. Jans, & R. Ghildyal (Eds.), Rhinoviruses: Methods and Protocols (Vol. 1221, pp. 171-180). (Methods in Molecular Biology). USA: Humana Press. https://doi.org/10.1007/978-1-4939-1571-2_13
WALKER, Erin ; Jensen, Joan ; GHILDYAL, Reena. / Reverse Genetic Engineering of the Human Rhinovirus Serotype 16 Genome to Introduce an Antibody-Detectable Tag. Rhinoviruses: Methods and Protocols. editor / D.A Jans ; R Ghildyal. Vol. 1221 USA : Humana Press, 2015. pp. 171-180 (Methods in Molecular Biology).
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WALKER, E, Jensen, J & GHILDYAL, R 2015, Reverse Genetic Engineering of the Human Rhinovirus Serotype 16 Genome to Introduce an Antibody-Detectable Tag. in DA Jans & R Ghildyal (eds), Rhinoviruses: Methods and Protocols. vol. 1221, Methods in Molecular Biology, Humana Press, USA, pp. 171-180. https://doi.org/10.1007/978-1-4939-1571-2_13

Reverse Genetic Engineering of the Human Rhinovirus Serotype 16 Genome to Introduce an Antibody-Detectable Tag. / WALKER, Erin; Jensen, Joan; GHILDYAL, Reena.

Rhinoviruses: Methods and Protocols. ed. / D.A Jans; R Ghildyal. Vol. 1221 USA : Humana Press, 2015. p. 171-180 (Methods in Molecular Biology).

Research output: A Conference proceeding or a Chapter in BookChapter

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WALKER E, Jensen J, GHILDYAL R. Reverse Genetic Engineering of the Human Rhinovirus Serotype 16 Genome to Introduce an Antibody-Detectable Tag. In Jans DA, Ghildyal R, editors, Rhinoviruses: Methods and Protocols. Vol. 1221. USA: Humana Press. 2015. p. 171-180. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-1571-2_13