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Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
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JoVE Journal Genetik
Reverse Genetics to Engineer Positive-Sense RNA Virus Variants

Reverse Genetics to Engineer Positive-Sense RNA Virus Variants

1,093 Views

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15:49 min

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June 09, 2022

DOI:

10.3791/63685-v

DOI:
10.3791/63685-v

•

15:49 min
June 09, 2022

•

3 Views
, , ,
1Virology Section, Department of Life Sciences,Shiv Nadar University, 2Gastroenterology,Sanjay Gandhi Postgraduate Institute of Medical Sciences

Summary

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The protocol describes a method for introducing controllable genetic diversity in the hepatitis C virus genome by combining full-length mutant RNA synthesis using error-prone PCR and reverse genetics. The method provides a model for phenotype selection and can be used for 10 kb long positive-sense RNA virus genomes.

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Reverse GeneticsPositive-sense RNA VirusViral GenomeMutagenesisRNA LibraryPhenotype SelectionCloning-freeEp-PCRIn Vitro RNA SynthesisCDNA SynthesisAmplificationA Overhang Addition

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Cite this Article

Soni, S., Agarwal, S., Aggarwal, R., Veerapu, N. S. Reverse Genetics to Engineer Positive-Sense RNA Virus Variants. J. Vis. Exp. (184), e63685, doi:10.3791/63685 (2022).

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