Rhinoviruses pp 171-180 | Cite as

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

  • Erin J. WalkerEmail author
  • Lora M. Jensen
  • Reena Ghildyal
Part of the Methods in Molecular Biology book series (MIMB, volume 1221)


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.

Key words

Human rhinovirus Reverse genetic engineering 



We thank Dr Wai-Ming Lee for the kind gift of anti-VP2 antibodies and the pR16.11 HRV16 plasmid.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

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

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