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Baculoviral Expression of Influenza A Virus (H1N1 New Caledonia) Neuraminidase in Insect Cells
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S Najafi   , F Behzadian * , F Fotouhi-Chahooki , M Tavasoti-Kheiri , J Fallah-Mehrabadi |
| Department of Molecular Genetics, Research Center for Sciences and Biotechnology, Malek Ashtar University, Tehran, Iran |
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Abstract: (5852 Views) |
| Background and Aims: Each year, the influenza virus causes moderate to severe infections with a high prevalence throughout the world. Accordingly, an influenza vaccine that ensures protection with only a single dose would be a much more cost effective approach to influenza prophylaxis. Generation of Influenza non-replicating virus-like particles (VLP) in baculoviral expression system is an attractive method for achieving this goal. One of the main components of such particles is Neuraminidase surficial glycoprotein that has important role to elicit humoral and cellular immune responses. Materials and Methods: In this study, the NA coding region amplified from the human influenza virus [A/New Caledonia 20/1999/ (H1N1)] was used to construct the NA recombinant bacmid into E.coli DH10Bac cells. Results: After verification of the new recombinant bacmid, it was transfected into the Spodoptera frugiperda (Sf9) insect cell line to generate recombinant baculovirus expressing the NA gene. The expression of NA in insect cells was confirmed by SDS-PAGE and western blot analysis. Conclusion: The recombinant baculovirus expressing the NA gene can be used in construction of influenza VLP when co-infect along with the other monocistronic baculoviruses expressing influenza Hemagglutinin and Matrix antigens. Moreover, the NA protein expressed in insect cells might be fully glycosylated and therefore is appropriate to be use in influenza vaccinology projects. |
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| Keywords: Influenza virus, SF9 cells, Baculovirus, Neuraminidase |
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Full-Text [PDF 324 kb]
(2308 Downloads)
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Type of Study: Original article |
Subject:
General
Received: 2014/11/7 | Accepted: 2014/11/7 | Published: 2014/11/7
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