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Expression of Influenza Heamagglutinin Globular Head in Different Eukaryotic Cells
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R Tavakoli   , F Fotouhi * , M Tavassoti-Kheiri , B Farahmand , M Saleh , M Tabatabaeiyan |
| Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran |
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Abstract: (5879 Views) |
| Background and Aims: Influenza (flu) is a respiratory infection in mammals and birds. It is caused by an RNA virus in the family Orthomyxoviridae. The virus is divided into three main types. Influenza virus type A is found in a wide variety of bird and mammal species and can undergo major shifts in immunological properties. Hemagglutinin (HA) is an important influenza virus surface antigen that is highly topical in influenza research. In the present study, the gene encoding HA1 protein which includes Hemagglutinin globular head from influenza virus A/Tehran/18/2010 (H1N1) was cloned into a eukaryotic expression plasmid (pCDNA3) and its expression was evaluated in eukaryotic cells. Materials and Methods: HA1 gene was incised from pFastBacTHc-HA1 by digestion, purified and subcloned into eukaryotic expression vector (pCDNA3). After verification of the cloning fidelity, the recombinant plasmid was transfected into COS-7 and BHK-21 cells, and its expression was detected by RT-PCR. Results: Restriction endonuclease digestion analysis, colony PCR and DNA sequencing indicated that the recombinant plasmid pCDNA3-HA1 had been constructed successfully. After transfection into eukaryotic cells, the presence of mRNA transcripts was verified by reverse transcriptase-polymerase chain reaction (RT-PCR). Conclusion: This study is a demonstrated success in the construction of eukaryotic expression plasmid for HA1 thus providing a basis for further probing into mechanism of virus infection and exploring DNA vaccine. |
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| Keywords: Influenza virus, Hemagglutinin, DNA vaccine |
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Full-Text [PDF 286 kb]
(1512 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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