Construction and expression of the eukaryotic expression vector for nucleoprotein gene of influenza virus A

Zhong Li; Li WangYi; Li MingYuan; Li Hong; Zhang WeiDong; Jiang ZhongHua

Chinese Journal of Zoonoses 23(4): 366-369

2007


Accession: 013235541

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Abstract
The eukaryotic expression vector of nucleoprotein(NP) gene on 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 2000 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.