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Related Concept Videos

Viral Recombination00:57

Viral Recombination

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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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Viral Mutations00:36

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A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material...
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The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
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Viral meningitis is the most common form of meningitis and is often referred to as aseptic meningitis to indicate the absence of bacterial involvement. It is generally milder than bacterial meningitis, with symptoms including fever, headache, stiff neck, drowsiness, nausea, photophobia, and vomiting. Rarely, more severe manifestations or death may occur. Common causative agents include enteroviruses, particularly coxsackie A and B viruses and echoviruses, all members of the Enterovirus genus...
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Arboviral Encephalitis01:25

Arboviral Encephalitis

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Arboviral encephalitis refers to brain inflammation caused by arthropod-borne viruses, particularly those transmitted through mosquito vectors. Among these, West Nile virus (WNV), a member of the Flaviviridae family, is a significant public health concern. WNV is an enveloped, positive-sense, single-stranded RNA virus. Human infection typically begins when an infected mosquito introduces the virus into the dermis during feeding. The primary transmission cycle involves birds as amplifying hosts...
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Encephalitis ll: Pathophysiology01:26

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Encephalitis is inflammation of the brain parenchyma caused by direct viral invasion or immune-mediated mechanisms triggered by infections or tumors. Both processes lead to neuronal injury, disrupted neurotransmission, and diverse neurological symptoms, often with overlapping clinical and pathological features.Autoimmune EncephalitisIn autoimmune encephalitis, antibodies target neuronal antigens on cell surfaces, synapses, or within neurons. A key example is anti-NMDAR encephalitis, which can...
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Related Experiment Video

Updated: May 5, 2026

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
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Reverse genetics system for Tick-borne encephalitis virus using Circular Polymerase Extension Reaction.

Saki Mitsunaga1, Tomokazu Tamura2, Samuel Nyampong1

  • 1Laboratory of Veterinary Microbiology, Joint Graduate School of Veterinary Medicine, Yamaguchi University, Yamaguchi, Japan.

Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases
|June 3, 2025
PubMed
Summary

Researchers successfully created infectious tick-borne encephalitis virus (TBEV) using an E. coli-free method called Circular Polymerase Extension Reaction (CPER). This new technique overcomes challenges in TBEV research and offers a valuable tool for studying this dangerous neurological pathogen.

Keywords:
Polymerase chain reaction-based Circular Polymerase Extension ReactionReverse genetics systemTick-borne encephalitis virus

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Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
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Area of Science:

  • Virology
  • Molecular Biology
  • Neuroscience

Background:

  • Tick-borne encephalitis virus (TBEV) is an Orthoflavivirus causing severe human neurological disease.
  • Traditional reverse genetics systems are hindered by the toxicity of Orthoflavivirus genomes to Escherichia coli.
  • An E. coli-free method, Circular Polymerase Extension Reaction (CPER), has shown promise for other RNA viruses.

Purpose of the Study:

  • To establish a recombinant TBEV production system using CPER.
  • To assess the viability and characteristics of TBEV generated via CPER.
  • To evaluate the infectivity and pathogenicity of CPER-derived TBEV in a mouse model.

Main Methods:

  • Application of the Circular Polymerase Extension Reaction (CPER) for TBEV genome assembly.
  • Genome sequencing and plaque-forming assays to characterize rescued TBEV strains (Oshima, Sofjin, Hypr).
  • Infectivity and pathogenicity studies in C57BL/6 mice.

Main Results:

  • Infectious recombinant TBEV strains (Oshima, Sofjin, Hypr) were successfully produced using CPER.
  • Rescued TBEVs exhibited cytopathic effects and efficient replication in BHK cells.
  • The CPER-derived TBEVs demonstrated intrinsic virulence in a mouse model, with observed nucleotide and amino acid substitutions.

Conclusions:

  • CPER is an effective E. coli-free method for generating infectious TBEV.
  • This approach facilitates the study of TBEV strains and holds potential for future research.
  • The successful rescue of TBEV via CPER provides a valuable tool for understanding flavivirus replication and pathogenesis.