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Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method
Background: Avian Influenza disease annually entails a significant economic loss to the poultry industry around the world. Influenza virus is a polymorphic virus of the orthomyxoviridae family (single-stranded RNA genome), and nucleoprotein (NP) is the structural and internal protein of the virus. T...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Iran University of Medical Sciences
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387826/ https://www.ncbi.nlm.nih.gov/pubmed/30815423 http://dx.doi.org/10.14196/mjiri.32.128 |
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author | Hosseini, Seyedeh Saeedeh Taheri, Kayvan Hosseini, Seyedeh Marzieh Gholami, Mehrdad Kouhsari, Ebrahim Edalati, Elahe Madani, Rasoul Mohammadzadeh, Rokhsare Zahedi Bialvaei, Abed Sholeh, Mohammad Golchin Far, Fariba |
author_facet | Hosseini, Seyedeh Saeedeh Taheri, Kayvan Hosseini, Seyedeh Marzieh Gholami, Mehrdad Kouhsari, Ebrahim Edalati, Elahe Madani, Rasoul Mohammadzadeh, Rokhsare Zahedi Bialvaei, Abed Sholeh, Mohammad Golchin Far, Fariba |
author_sort | Hosseini, Seyedeh Saeedeh |
collection | PubMed |
description | Background: Avian Influenza disease annually entails a significant economic loss to the poultry industry around the world. Influenza virus is a polymorphic virus of the orthomyxoviridae family (single-stranded RNA genome), and nucleoprotein (NP) is the structural and internal protein of the virus. The aim of the work was to purify nucleoprotein for further investigations with a simple, low-cost, fast and practical method. Methods: In this study, H9N2 influenza virus was isolated in specific pathogen-free embryonated chicken eggs by allantoically inoculating 103 to 105 egg-infective doses (EID50) for 9 to 11 days, purified by 10% (W/V) polyethylene glycol (PEG) 6000 with a sucrose gradient of 60% to 30%. The influenza virus proteins were collected and prepared as fractions by preparative electrophoresis. Finally, the purified NP was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot procedures. Results: The protein analysis with SDS-PAGE and silver nitrate staining indicated that the desired samples contained purified nucleoprotein and lacked other viral proteins. The results of the investigation of lyophilized fractions containing nucleoprotein on the SDS-PAGE revealed the absence of viral RNA in nucleoprotein and its high purity. Conclusion: According to this study, purified nucleoprotein can be used to produce nucleoprotein vaccines, as well as to study structural, molecular and diagnostic and therapeutic materials. |
format | Online Article Text |
id | pubmed-6387826 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Iran University of Medical Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-63878262019-02-27 Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method Hosseini, Seyedeh Saeedeh Taheri, Kayvan Hosseini, Seyedeh Marzieh Gholami, Mehrdad Kouhsari, Ebrahim Edalati, Elahe Madani, Rasoul Mohammadzadeh, Rokhsare Zahedi Bialvaei, Abed Sholeh, Mohammad Golchin Far, Fariba Med J Islam Repub Iran Original Article Background: Avian Influenza disease annually entails a significant economic loss to the poultry industry around the world. Influenza virus is a polymorphic virus of the orthomyxoviridae family (single-stranded RNA genome), and nucleoprotein (NP) is the structural and internal protein of the virus. The aim of the work was to purify nucleoprotein for further investigations with a simple, low-cost, fast and practical method. Methods: In this study, H9N2 influenza virus was isolated in specific pathogen-free embryonated chicken eggs by allantoically inoculating 103 to 105 egg-infective doses (EID50) for 9 to 11 days, purified by 10% (W/V) polyethylene glycol (PEG) 6000 with a sucrose gradient of 60% to 30%. The influenza virus proteins were collected and prepared as fractions by preparative electrophoresis. Finally, the purified NP was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot procedures. Results: The protein analysis with SDS-PAGE and silver nitrate staining indicated that the desired samples contained purified nucleoprotein and lacked other viral proteins. The results of the investigation of lyophilized fractions containing nucleoprotein on the SDS-PAGE revealed the absence of viral RNA in nucleoprotein and its high purity. Conclusion: According to this study, purified nucleoprotein can be used to produce nucleoprotein vaccines, as well as to study structural, molecular and diagnostic and therapeutic materials. Iran University of Medical Sciences 2018-12-21 /pmc/articles/PMC6387826/ /pubmed/30815423 http://dx.doi.org/10.14196/mjiri.32.128 Text en © 2018 Iran University of Medical Sciences http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial 3.0 License (CC BY-NC 3.0), which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly. |
spellingShingle | Original Article Hosseini, Seyedeh Saeedeh Taheri, Kayvan Hosseini, Seyedeh Marzieh Gholami, Mehrdad Kouhsari, Ebrahim Edalati, Elahe Madani, Rasoul Mohammadzadeh, Rokhsare Zahedi Bialvaei, Abed Sholeh, Mohammad Golchin Far, Fariba Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method |
title | Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method |
title_full | Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method |
title_fullStr | Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method |
title_full_unstemmed | Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method |
title_short | Extraction and purification of the H9N2 virus nucleoprotein: A simple and practical method |
title_sort | extraction and purification of the h9n2 virus nucleoprotein: a simple and practical method |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387826/ https://www.ncbi.nlm.nih.gov/pubmed/30815423 http://dx.doi.org/10.14196/mjiri.32.128 |
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