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Updated: May 19, 2026
Multi-target Parallel Processing Approach for Gene-to-structure Determination of the Influenza Polymerase PB2 Subunit
Published on: June 28, 2013
[Comparison of three methods in construction fusion gene of influenza A virus Nucleoprotein]
Bo Liu1, Bao-Lin Wang, Ling Chen
1Department of Respiratory Medicine, Laboratory of Respiratory Medicine Research, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China.
Objective:
To develop a best method of constructing influenza NP fusion gene containing enhanced green fluorescent protein (EGFP).
Methods:
The full-length NP gene of influenza A was amplified by RT-PCR and was inserted into an eukaryotic expression vector pEGFP-N1 in order to construct a fusion gene of pEGFP-N1-NP using three different methods. Method one, NP gene containing restriction endonucleases and pEGFP-N1 were both digested using the same restrict enzymes and ligated, yielding the fusion gene of pEGFP-N1-NP. Method two, NP gene was cloned into pMD19-T Vector to construct a plasmid of pMD19-T-NP. The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP; Method three, NP gene containing restriction endonucleases was cloned into pMD19-T Simple Vector to construct a plasmid of pMD19-T-NP. The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP.
Results:
The fusion gene of recombinant eukaryotic expression vector pEGFP-N1-NP was successfully constructed by using method three.
Conclusions:
The full-length NP gene is obtained and its fusion gene of recombinant eukaryotic expression plasmid is successfully constructed. This study provides foundation for further understanding the biological function of NP protein and the mechanism of diseases induced by influenza A virus.
