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Related Concept Videos

Viral Recombination00:57

Viral Recombination

23.5K
Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
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Conservative Site-specific Recombination and Phase Variation02:53

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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PCR01:32

PCR

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Overview
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Related Experiment Video

Updated: Jul 20, 2025

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection
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Rapid Generation of Recombinant Flaviviruses Using Circular Polymerase Extension Reaction.

Hao-Long Dong1, Mei-Juan He1, Qing-Yang Wang1

  • 1Academy of Military Medical Sciences, Beijing 100071, China.

Vaccines
|July 29, 2023
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Summary

A novel Circular Polymerase Extension Reaction (CPER) method rapidly generates recombinant flaviviruses, including Japanese encephalitis virus (JEV) and Zika virus (ZIKV), in both insect and vertebrate cells. This bacterium-free approach simplifies flavivirus research and aids in developing future outbreak countermeasures.

Keywords:
CPERFlavivirusreporter virusreverse geneticvaccine

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Area of Science:

  • Virology
  • Molecular Biology
  • Biotechnology

Background:

  • Flaviviruses are significant arthropod-borne pathogens impacting human and animal health.
  • Conventional flavivirus reverse genetics is hindered by genome toxicity in bacteria, complicating clone construction.
  • Existing methods for generating infectious flavivirus clones are often time-consuming and labor-intensive.

Purpose of the Study:

  • To develop a rapid, efficient, and bacterium-free method for flavivirus reverse genetics.
  • To establish a versatile platform for synthesizing recombinant flaviviruses in diverse cell types.
  • To facilitate the creation of novel flavivirus constructs for research and countermeasure development.

Main Methods:

  • Application of a Circular Polymerase Extension Reaction (CPER) for de novo synthesis of flavivirus genomes.
  • Recovery of recombinant Japanese encephalitis virus (JEV) vaccine strain SA-14-14-2 in Vero cells.
  • Adaptation of the CPER method for rescuing insect-specific flaviviruses (ISFs) in C6/36 mosquito cells.
  • Generation of chimeric Zika virus (ChinZIKV) and a Culex flavivirus (CxFV) reporter virus (CxFV-GFP) using CPER.

Main Results:

  • Successful de novo synthesis and recovery of JEV SA-14-14-2 using CPER in vertebrate cells.
  • Efficient rescue of ISFs, including ChinZIKV and CxFV-GFP, in insect cells via modified CPER.
  • Demonstration of CPER as a rapid and bacterium-free approach for flavivirus generation.
  • Establishment of a versatile CPER-based system for flaviviruses with different host ranges.

Conclusions:

  • CPER provides a simple and rapid method for generating diverse flaviviruses and potentially other RNA viruses.
  • The established CPER-based recovery system streamlines flavivirus research across different host ranges.
  • This methodology holds promise for accelerating the development of effective countermeasures against emerging flavivirus threats.