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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 19, No. 1, 2016, pp. 61-64
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Bioline Code: ej16010
Full paper language: English
Document type: Short Communication
Document available free of charge
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Electronic Journal of Biotechnology, Vol. 19, No. 1, 2016, pp. 61-64
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Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
Kanehira, Katsushi; Uchida, Yuko & Saito, Takehiko
Abstract
Background:
Avian influenza viruses (AIVs) are influenza A viruses which are isolated from domestic and wild
birds. AIVs that include highly pathogenic avian influenza viruses (HPAIVs) are a major concern to the poultry
industry because they cause outbreaks in poultry with extraordinarily high lethality. In addition, AIVs threaten
human health by occasional zoonotic infection of humans from birds. Tools to visualize AIV-infected cells
would facilitate the development of diagnostic tests and preventative methods to reduce the spread of AIVs. In
this study, a self-assembling split-green fluorescent protein (split-GFP) system, combined with influenza virus
reverse genetics was used to construct a visualization method for influenza virus-infected cells.
Results:
The viral nucleoprotein (NP) segment of AIV was genetically modified to co-express GFP11 of
self-assembling split-GFP, and the recombinant AIV with the modified NP segment was generated by
plasmid-based reverse genetics. Infection with the recombinant AIV in cultured chicken cells was visualized by
transient transfection with a GFP1-10 expression vector and fluorescence was observed in the cells at 96 hours
post-inoculation. Virus titer of the recombinant AIV in embryonated eggs was comparable to wild type AIV
titers at 48 h post inoculation. The inserted sequence encoding GFP11 was stable for up to ten passages in
embryonated eggs.
Conclusions:
A visualization system for AIV-infected cells using split-GFP was developed. This method could be
used to understand AIV infection dynamics in cells.
Keywords
Avian influenza virus; Reverse genetics; Split-GFP
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