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Rhinoviruses pp 149-170 | Cite as

Reverse Genetics System for Studying Human Rhinovirus Infections

  • Wai-Ming LeeEmail author
  • Wensheng Wang
  • Yury A. Bochkov
  • Iris Lee
Protocol
  • 1.8k Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 1221)

Abstract

Human rhinovirus (HRV) contains a 7.2 kb messenger-sense RNA genome which is the template for reproducing progeny viruses after it enters the cytoplasm of a host cell. Reverse genetics refers to the regeneration of progeny viruses from an artificial cDNA copy of the RNA genome of an RNA virus. It has been a powerful molecular genetic tool for studying HRV and other RNA viruses because the artificial DNA stage makes it practical to introduce specific mutations into the viral RNA genome. This chapter uses HRV-16 as the model virus to illustrate the strategy and methods for constructing and cloning the artificial cDNA copy of a full-length HRV genome, identifying the infectious cDNA clone isolates, and selecting the most vigorous cDNA clone isolate to serve as the standard parental clone for future molecular genetic study of the virus.

Key words

RNA virus Full-length infectious cDNA clone In vitro transcription Transfection 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Wai-Ming Lee
    • 1
    Email author
  • Wensheng Wang
    • 2
  • Yury A. Bochkov
    • 3
  • Iris Lee
    • 1
  1. 1.Biological Mimetics Inc.FrederickUSA
  2. 2.Global Biological DevelopmentBayer HealthcareBerkeleyUSA
  3. 3.Department of Pediatrics, School of Medicine and Public HealthUniversity of WisconsinMadisonUSA

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