Construction of a bicistronic eukaryotic vector expressing M۱ and NP genes of influenza virus

Objective: In this study, two conserved genes (M۱ and NP) of influenza virus were expressed in a bicistronic vector in order to develop a universal gene based vaccine. Materials and Methods: Plasmids M۱-pIRES۲-EGFP, pIRES۲-NP were constructed by cloning the PCR products of M۱ and NP genes which were amplified from the A/Peurto Rico/۸/۳۴ (H۱N۱) influenza virus strain into the plasmid expression vector pIRES۲-EGFP, respectively. For construction of M۱-pIRES۲-NP bicistronic plasmid, M۱ gene was extracted from M۱-pIRES-EGFP plasmid and sub-cloned into pIRES۲-NP construct. Finally, simultaneous expression of both genes was assessed by transient transfection of bicistronic plasmid into BHK-۲۱ 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 M۱-pIRES۲-NP plasmid expression in BHK-۲۱ cell lines confirmed by immunofluoresence microscopy. Conclusion: Simultaneous expression of influenza M۱ and NP genes from a bicistronic plasmid containing “IRES” sequence is achievable.

کلیدواژه ها:

Influenza Virus ، bicistronic vector ، Simultaneous Expression ، M۱ ، NP ، ژن نوکلئوپروتئین ، ویروس آنفلوانزا ، ژن M۱ ، بیان همزمان ، ناقل دو سیسترونی