Abstract:The eukaryotic expression vector for nucleoprotein(NP) gene of influenza A virus A/PR/8/34(H1N1)was constructed by transfecting this gene into HEK293 cells and detecting the expression of the target protein. The NP gene was cloned by RT-PCR and then inserted into the eukaryotic expression vector pcDNA3.1(+).After identification with restriction enzyme digestion,PCR assay and sequencing,the recombinant plasmid was transfected into HEK293 cells with lipofectamineTM 2 000 induction. The transient expression of target protein was observed by immunofluorescence assay. It was found that the NP gene could be amplified by using the RT-PCR and the expression of the target protein could be demonstrated through the detection with immunofluorescence assay after insertion into the eukaryotic expression vector pcDNA3.1(+) and transfection to HEK293 cells. It is evident that the eukaryotic expression vector for parts of NP gene sequence of influenza virus A was successfully constructed for use of further studies on nucleoprotein and for effective DNA vaccine of influenza virus A.
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