Shenagari, Mohammad, Sabahi, Farzaneh, Tavasoti Kheiri, Masoumah, Forghan Parast, Kambiz, Jamali, Abbas, Hashemi, Hamidreza, Khodamoradi, Shadi (2011). Construction of a bicistronic eukaryotic vector expressing M1 and NP genes of influenza virus. , 14(2), 51-62.
Mohammad Shenagari; Farzaneh Sabahi; Masoumah Tavasoti Kheiri; Kambiz Forghan Parast; Abbas Jamali; Hamidreza Hashemi; Shadi Khodamoradi. "Construction of a bicistronic eukaryotic vector expressing M1 and NP genes of influenza virus". , 14, 2, 2011, 51-62.
Shenagari, Mohammad, Sabahi, Farzaneh, Tavasoti Kheiri, Masoumah, Forghan Parast, Kambiz, Jamali, Abbas, Hashemi, Hamidreza, Khodamoradi, Shadi (2011). 'Construction of a bicistronic eukaryotic vector expressing M1 and NP genes of influenza virus', , 14(2), pp. 51-62.
Shenagari, Mohammad, Sabahi, Farzaneh, Tavasoti Kheiri, Masoumah, Forghan Parast, Kambiz, Jamali, Abbas, Hashemi, Hamidreza, Khodamoradi, Shadi Construction of a bicistronic eukaryotic vector expressing M1 and NP genes of influenza virus. , 2011; 14(2): 51-62.

Construction of a bicistronic eukaryotic vector expressing M1 and NP genes of influenza virus

Pathobiology Reserach
Article 5, Volume 14, Issue 2, 2011, Pages 51-62    PDF (869.38 K)
Authors
Mohammad Shenagari1; Farzaneh Sabahi* 2; Masoumah Tavasoti Kheiri3; Kambiz Forghan Parast4; Abbas Jamali5; Hamidreza Hashemi6; Shadi Khodamoradi7
1Ph.D. Student, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University
2Associated Professor, Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University
3Assistant Professor, Department of Influenza, Pasteur Institute of Iran
4Associated Professor, Department of Bacteriology, Faculty of Medicine, Gilan University of Medical Sciences, Iran
5Post-Doctoral fellow, Department of Influenza, Pasteur Institute of Iran, Iran
6Ph.D. Student, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Iran
7M.Sc. Student, Department of Influenza, Pasteur Institute of Iran, Iran
Abstract
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.
Keywords
Influenza Virus; bicistronic vector; Simultaneous Expression; M1; nP
Statistics
Article View: 106
PDF Download: 53
Statistics
Number of Journals45
Number of Issues2,160
Number of Articles24,584
Article View20,343,939
PDF Download16,248,656