Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: Jul 7, 2026

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection
09:25

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection

Published on: May 24, 2020

A PCR-based protocol for generating West Nile virus replicons.

Junko Maeda1, Hirotaka Takagi, Shingo Hashimoto

  • 1Department of Prion Diseases, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.

Journal of Virological Methods
|February 5, 2008
PubMed
Summary

A novel protocol efficiently generates West Nile virus (WNV) replicons in one week. This method enables rapid synthesis of stable WNV replicon cDNAs for research applications.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

NUP107-160 nuclear pore sub-complex members determine symbiotic ion channel localization in legumes.

Plant physiology·2026
Same author

A microfluidic spore chamber for long-term imaging of single-spore hyphal development.

Fungal genetics and biology : FG & B·2026
Same author

A Rare Case of Adult-Onset Human Parechovirus Type A3 Encephalitis.

Internal medicine (Tokyo, Japan)·2026
Same author

Propagation of human sapovirus GII.7 strain in HuTu80 subcloned cells.

Japanese journal of infectious diseases·2026
Same author

Differential Roles of BRCA1 and BRCA2 in the DNA Damage Response Revealed by isogenic V79 Mutant Cell lines.

Mutagenesis·2026
Same author

Generation of NBS1 knockout in Chinese hamster cells revealed ATR role for radiation and etoposide induced DNA damage in absence of NBS1 proteins.

Frontiers in oncology·2026

Area of Science:

  • Virology
  • Molecular Biology
  • Biotechnology

Background:

  • West Nile virus (WNV) poses a significant public health concern.
  • Efficient generation of WNV replicons is crucial for studying viral replication and developing countermeasures.

Purpose of the Study:

  • To develop a novel and rapid protocol for the synthesis of West Nile virus (WNV) replicons.
  • To enable efficient generation of stable WNV replicon cDNAs for downstream applications.

Main Methods:

  • Utilized polymerase chain reactions (PCRs) to amplify fragmented cDNAs covering the entire WNV RNA sequence.
  • Employed overlapping sequences for fragment annealing and DNA polymerase elongation.
  • Generated full-sized WNV replicon cDNAs through short-cycle PCRs.
  • Transcribed WNV replicons in vitro using synthesized cDNAs.

More Related Videos

Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone
07:35

Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone

Published on: June 7, 2017

High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication
10:16

High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication

Published on: October 30, 2021

Related Experiment Videos

Last Updated: Jul 7, 2026

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection
09:25

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection

Published on: May 24, 2020

Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone
07:35

Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone

Published on: June 7, 2017

High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication
10:16

High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication

Published on: October 30, 2021

Main Results:

  • Successfully generated full-sized WNV replicon cDNAs.
  • In vitro-transcribed WNV replicons were introduced into mammalian cells.
  • Detected viral envelope protein and both positive- and negative-strand viral RNAs in transfected cells.
  • Demonstrated the stability of the synthetic replicon sequence using high-fidelity DNA polymerase.

Conclusions:

  • The developed protocol provides a rapid (1-week) method for WNV replicon synthesis.
  • The protocol yields stable WNV replicons suitable for research.
  • This advancement facilitates WNV research and the development of antiviral strategies.