Reverse genetics system for studying human rhinovirus infections

Wai Ming Lee; Wensheng Wang; Yury A. Bochkov; Iris Lee

doi:10.1007/978-1-4939-1571-2_12

Reverse genetics system for studying human rhinovirus infections

Wai Ming Lee, Wensheng Wang, Yury A. Bochkov, Iris Lee

Division of Rheumatology

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

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.

Original language	English
Pages (from-to)	149-170
Number of pages	22
Journal	Methods in Molecular Biology
Volume	1221
DOIs	https://doi.org/10.1007/978-1-4939-1571-2_12
State	Published - 2015

Keywords

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

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10.1007/978-1-4939-1571-2_12

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AB - 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.

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Reverse genetics system for studying human rhinovirus infections

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