Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein

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 letha...

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Autores principales:	Kanehira,Katsushi, Uchida,Yuko, Saito,Takehiko
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2016
Materias:	Avian influenza virus Reverse genetics Split-GFP
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000100010
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spelling	oai:scielo:S0717-345820160001000102016-05-11Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent proteinKanehira,KatsushiUchida,YukoSaito,Takehiko Avian influenza virus Reverse genetics Split-GFP 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.19 n.1 20162016-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000100010en10.1016/j.ejbt.2015.08.008
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Avian influenza virus Reverse genetics Split-GFP
spellingShingle	Avian influenza virus Reverse genetics Split-GFP Kanehira,Katsushi Uchida,Yuko Saito,Takehiko Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
description	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.
author	Kanehira,Katsushi Uchida,Yuko Saito,Takehiko
author_facet	Kanehira,Katsushi Uchida,Yuko Saito,Takehiko
author_sort	Kanehira,Katsushi
title	Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
title_short	Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
title_full	Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
title_fullStr	Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
title_full_unstemmed	Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
title_sort	visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2016
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000100010
work_keys_str_mv	AT kanehirakatsushi visualizationofavianinfluenzavirusinfectedcellsusingselfassemblingfragmentsofgreenfluorescentprotein AT uchidayuko visualizationofavianinfluenzavirusinfectedcellsusingselfassemblingfragmentsofgreenfluorescentprotein AT saitotakehiko visualizationofavianinfluenzavirusinfectedcellsusingselfassemblingfragmentsofgreenfluorescentprotein
_version_	1718441925120360448

Visualization of avian influenza virus infected cells using self-assembling fragments of green fluorescent protein

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