Volume 14, Issue 2 (2011)
mjms 2011, 14(2): 51-62 |
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Shenagari M, Sabahi F, Tavasoti Kheiri M, Forghan Parast K, Jamali A, Hashemi H et al . Construction of a bicistronic eukaryotic vector expressing M1 and NP genes of influenza virus. mjms 2011; 14 (2) :51-62
URL: http://mjms.modares.ac.ir/article-30-8566-en.html
URL: http://mjms.modares.ac.ir/article-30-8566-en.html
Mohammad Shenagari1 
, Farzaneh Sabahi 2, Masoumah Tavasoti Kheiri3 , Kambiz Forghan Parast4 , Abbas Jamali5 , Hamidreza Hashemi6 , Shadi Khodamoradi7
, Farzaneh Sabahi 2, Masoumah Tavasoti Kheiri3 , Kambiz Forghan Parast4 , Abbas Jamali5 , Hamidreza Hashemi6 , Shadi Khodamoradi7
1- Ph.D. Student, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University
2- Associated Professor, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University
3- Assistant Professor, Department of Influenza, Pasteur Institute of Iran
4- Associated Professor, Department of Bacteriology, Faculty of Medicine, Gilan University of Medical Sciences, Iran
5- Post-Doctoral fellow, Department of Influenza, Pasteur Institute of Iran, Iran
6- Ph.D. Student, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Iran
7- M.Sc. Student, Department of Influenza, Pasteur Institute of Iran, Iran
2- Associated Professor, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University
3- Assistant Professor, Department of Influenza, Pasteur Institute of Iran
4- Associated Professor, Department of Bacteriology, Faculty of Medicine, Gilan University of Medical Sciences, Iran
5- Post-Doctoral fellow, Department of Influenza, Pasteur Institute of Iran, Iran
6- Ph.D. Student, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Iran
7- M.Sc. Student, Department of Influenza, Pasteur Institute of Iran, Iran
Abstract: (5198 Views)
Objective: In this study, two conserved genes (M1 and NP) of influenza virus were expressed in a bicistronic vector in order to develop a universal gene based vaccine.
Materials and Methods: Plasmids M1-pIRES2-EGFP, pIRES2-NP were constructed by cloning the PCR products of M1 and NP genes which were amplified from the A/Peurto Rico/8/34 (H1N1) influenza virus strain into the plasmid expression vector pIRES2-EGFP, respectively. For construction of M1-pIRES2-NP bicistronic plasmid, M1 gene was extracted from M1-pIRES-EGFP plasmid and sub-cloned into pIRES2-NP construct. Finally, simultaneous expression of both genes was assessed by transient transfection of bicistronic plasmid into BHK-21 cell lines and subsequent immunofluorescence staining.
Results: The results of enzymatic double digestions on the constructed plasmids and sequencing demonstrated the success of cloning processes of above mentioned genes. Correct expression of these genes was confirmed by M1-pIRES2-NP plasmid expression in BHK-21 cell lines confirmed by immunofluoresence microscopy.
Conclusion: Simultaneous expression of influenza M1 and NP genes from a bicistronic plasmid containing “IRES” sequence is achievable.
Article Type: Original Manuscipt |
Subject:
Molecular|Virology
Received: 2011/03/15 | Accepted: 2011/06/11
Received: 2011/03/15 | Accepted: 2011/06/11
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